JP2024527517A - Anti-PD1 antibody preparation - Google Patents

Abstract

The present invention relates to an anti-PD1 antibody pharmaceutical composition comprising a citrate buffer, one or more amino acids other than histidine, and a surfactant.

Description

The present invention relates to an anti-PD1 antibody pharmaceutical composition containing a citrate buffer, one or more amino acids other than histidine, and a surfactant.

Programmed cell death protein 1 (PD-1) is a cell surface protein that acts as an immune checkpoint and prevents autoimmune diseases by suppressing T-cell inflammatory activity. PD-1 binds to two ligands, PD-L1 and PD-L2. PD-L1 overexpression has been found in various types of cancer, and it has been found that inhibition of the interaction between PD-1 and PD-L1 can enhance T-cell responses, thereby mediating antitumor activity.

Several antibodies have been developed that target either PD-1 or PD-L1. Pembrolizumab, marketed under the trade name Keytruda®, is a humanized IgG4 antibody that binds to PD-1 and blocks its interaction with PD-L1. It is currently approved for the treatment of several types of cancer, including melanoma, non-small cell lung cancer, squamous cell carcinoma of the head and neck, bladder cancer, and Hodgkin's lymphoma. Nivolumab, marketed under the trade name Opdivo®, is a fully human IgG4 antibody that binds to PD-1 and blocks its interaction with PD-L1. It is currently approved for the treatment of melanoma, non-small cell lung cancer, and renal cell carcinoma.

US Patent No. 5,993,333 and US Patent No. 5,993,333 disclose liquid and lyophilized formulations of pembrolizumab containing a histidine buffer, polysorbate 80 and sucrose.
Patent Document 3 discloses an anti-PD1 antibody formulation containing sodium acetate, α,α-trehalose dihydrate and polysorbate 20, having a pH of 5.2.

Patent document 4 describes a pharmaceutical formulation containing an anti-PD1 antibody, citrate, histidine, mannitol, sodium chloride, edetate, and polysorbate 20 or polysorbate 80, and having a pH of 5.5 to 6.5.

Patent Document 5 discloses a pharmaceutical composition comprising an anti-PD1 antibody, histidine, sucrose, proline and polysorbate 80, at a pH of 6.0.
US Patent No. 6,399,633 describes an anti-PD1 antibody formulation containing a buffer, a stabilizer, a non-ionic surfactant and an antioxidant.

Patent Document 7 discloses a pharmaceutical composition comprising an anti-PD1 antibody, histidine, sucrose, polysorbate 20 and EDTA, at a pH of 6.5.
Patent document 8 describes a pharmaceutical composition comprising an anti-PD1 antibody, a disaccharide, a buffer, a chelating agent and a polysorbate, the composition having a pH of 4.5 to 5.5.

US Patent No. 5,999,333 describes a pharmaceutical composition comprising an anti-PD1 antibody, a buffer, a stabilizer, a surfactant and an antioxidant.
Patent Document 10 discloses a buffer-free anti-PD1 antibody pharmaceutical formulation.

US Patent No. 5,999,943 describes a liquid pharmaceutical composition comprising an anti-PD1 antibody, a histidine or citrate buffer, a sugar or sugar alcohol, and a non-ionic surfactant.
Nonetheless, there remains a need for stable anti-PD1 antibody formulations that are safe for administration to human patients.

International Publication No. 2012/135408 International Publication No. 2019/160751 International Publication No. 2017/054646 International Publication No. 2018/028383 International Publication No. 2018/187057 International Publication No. 2018/204368 International Publication No. 2019/142149 International Publication No. 2019/171253 International Publication No. 2020/097141 International Publication No. 2021/118321 International Publication No. 2021/123202

The present invention also relates to a liquid pharmaceutical composition comprising: (a) an anti-human PD1 antibody; (b) one or more amino acids other than histidine; (c) a non-ionic surfactant; and (d) a citrate buffer, the pH of which is 5.0 to 5.8, preferably 5.5.

In one embodiment, the citrate buffer is present at a concentration of 1 mM to 50 mM, preferably at a concentration of 5 mM or 10 mM.
In one embodiment, the non-ionic surfactant is polysorbate 20 or polysorbate 80, preferably polysorbate 80.

In one embodiment, the non-ionic surfactant is present in a concentration of from 0.1 mg/ml to 0.4 mg/ml, preferably at a concentration of 0.2 mg/ml.
In one embodiment, the amino acids are arginine and/or lysine.

In one embodiment, the pharmaceutical composition comprises arginine at a concentration of 100 mM to 350 mM, or at a concentration of 250 mM to 350 mM.
In one embodiment, the pharmaceutical composition comprises lysine at a concentration of 100 mM to 300 mM.

In one embodiment, the pharmaceutical composition comprises arginine and lysine, wherein the total concentration of arginine and lysine is between 100 mM and 350 mM.
In one embodiment, the anti-human PD1 antibody is pembrolizumab.

In one embodiment, the anti-human PD1 antibody is present at a concentration of 25-80 mg/ml.
The present invention also relates to a liquid pharmaceutical composition consisting of citrate buffer, arginine and/or lysine, polysorbate 20 or polysorbate 80, pembrolizumab and water for injection, and having a pH of 5.0 to 5.8.

In one embodiment, the liquid pharmaceutical composition comprises 5 or 10 mM citrate buffer, 300 mM arginine, 0.2 mg/ml polysorbate 80, 25 mg/ml pembrolizumab, and water for injection, and has a pH of 5.0 to 5.8.

In one embodiment, the liquid pharmaceutical composition comprises 10 mM citrate buffer, 250 mM lysine, 0.2 mg/ml polysorbate 80, 25 mg/ml pembrolizumab, and water for injection, and has a pH of 5.0-5.8.

In one embodiment, the liquid pharmaceutical composition comprises 10 mM citrate buffer, 150 mM arginine, 150 mM lysine, 0.2 mg/ml polysorbate 80, 25 mg/ml pembrolizumab, and water for injection, and has a pH of 5.0-5.8.

The present invention also relates to a liquid pharmaceutical composition comprising 5 mM citrate buffer, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80, 25 mg/ml pembrolizumab and water for injection, and having a pH of 5.0 to 5.8, preferably a pH of 5.5.

The present invention also relates to a liquid pharmaceutical composition comprising L-histidine/histidine hydrochloride, sucrose or trehalose dihydrate, polysorbate 20, 90-350 mg/ml pembrolizumab and water for injection, and having a pH of 5.2-5.8.

In one embodiment, the liquid pharmaceutical composition comprises 10 mM L-histidine/histidine hydrochloride, 140-220 mM trehalose dihydrate, 0.1 mg/ml polysorbate 20, 100 or 150 mg/ml pembrolizumab, and water for injection, and has a pH of 5.2-5.8.

The present invention also relates to a liquid pharmaceutical composition comprising citrate buffer, sucrose or trehalose dihydrate, polysorbate 80, 90-350 mg/ml pembrolizumab, and water for injection, and having a pH of 5.2-5.8.

The present invention also relates to a liquid pharmaceutical composition comprising 10 mM citrate buffer, 140-220 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80, 100 mg/ml or 150 mg/ml pembrolizumab, and water for injection, and having a pH of 5.0-5.8.

The present invention also relates to a liquid pharmaceutical composition comprising 10 mM citrate buffer, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80, 100 mg/ml or 150 mg/ml pembrolizumab, and water for injection, and having a pH of 5.0 to 5.8.

In one embodiment, the liquid pharmaceutical composition is for use in the treatment of cancer.
In one embodiment, the cancer can be used to treat melanoma, non-small cell lung cancer, classical Hodgkin's lymphoma, urothelial carcinoma, squamous cell carcinoma of the head and neck, renal cell carcinoma, colorectal carcinoma, small cell lung cancer, microsatellite instability high or mismatch repair deficient cancer, primary mediastinal large B cell lymphoma, gastric or gastroesophageal junction adenocarcinoma, hepatocellular carcinoma, Merkel cell carcinoma, endometrial cancer, oncogene hypermutated cancer, cutaneous squamous cell carcinoma, triple negative breast cancer or cervical cancer.

FIG. 1 is a contour plot generated by DOE software showing SE-HPLC predicted HMWS peak delta (after 2 weeks storage at 40° C.) as a function of citric acid monohydrate concentration and pH parameters. FIG. 1 is a contour plot generated by DOE software showing SE-HPLC predicted HMWS peak delta (after 2 weeks storage at 40° C.) as a function of factors pH and concentration of trehalose dihydrate. FIG. 13 is a contour plot generated by DOE software showing the predicted SE-HPLC monomer peak delta (after 2 weeks storage at 40° C.) as a function of citric acid monohydrate concentration and pH factors. FIG. 1 is a contour plot generated by DOE software showing the SE-HPLC predicted LMWS peak delta (after 2 weeks storage at 40° C.) as a function of factors pH and concentration of trehalose dihydrate. FIG. 1 is a contour plot generated by DOE software showing SE-HPLC predicted LMWS peak delta (after 2 weeks storage at 40° C.) as a function of citric acid monohydrate concentration and pH parameters. FIG. 1 is a contour plot generated by DOE software showing SE-HPLC predicted LMWS peak delta (after 2 weeks storage at 40° C.) as a function of factors of trehalose dihydrate concentration and polysorbate 20 (PS20) concentration. FIG. 1 is a contour plot generated by DOE software showing CEX-HPLC predicted acidic peak delta (after 2 weeks storage at 40° C.) as a function of citric acid monohydrate concentration and pH factors. FIG. 1 is a contour plot generated by DOE software showing CEX-HPLC predicted acidic peak delta (after 2 weeks storage at 40° C.) as a function of the factors citric acid monohydrate concentration and trehalose dihydrate concentration. FIG. 1 is a contour plot generated by DOE software showing the CEX-HPLC predicted main peak delta (after 2 weeks storage at 40° C.) as a function of citric acid monohydrate concentration and pH factors. FIG. 1 is a contour plot generated by DOE software showing the CEX-HPLC predicted main peak delta (after 2 weeks storage at 40° C.) as a function of the parameters pH and concentration of trehalose dihydrate. FIG. 1 is a contour plot generated by DOE software showing SE-HPLC predicted HMWS peak delta (after 2 weeks storage at 40° C.) as a function of factors of lysine HCl concentration and arginine HCl concentration. FIG. 13 is a contour plot generated by DOE software showing the predicted SE-HPLC monomer peak delta (after 2 weeks storage at 40° C.) as a function of the factors Lysine HCl concentration and Arginine HCl concentration. FIG. 1 is a contour plot generated by DOE software showing CEX-HPLC predicted acidic peak delta (after 2 weeks storage at 40° C.) as a function of factors of Lysine HCl concentration and Arginine HCl concentration. FIG. 1 is a contour plot generated by DOE software showing CEX-HPLC predicted acidic peak delta (after 2 weeks storage at 40° C.) as a function of the factors arginine HCl concentration and glycerol concentration. FIG. 1 is a contour plot generated by DOE software showing the CEX-HPLC predicted main peak delta (after 2 weeks storage at 40° C.) as a function of factors of lysine HCl concentration and glycerol concentration. FIG. 13 is a contour plot generated by the DOE software showing the CEX-HPLC predicted main peak delta (after 2 weeks storage at 40° C.) as a function of the factors arginine HCl concentration and glycerol concentration.

The present invention, as illustratively described below, may suitably be practiced in the absence of any element or elements, limitation or limitations not specifically disclosed herein.
The present invention will be described in terms of particular embodiments but the invention is not limited thereto but only by the claims.

When the term "comprising" is used in the present specification and claims, it does not exclude other elements. For the purposes of the present invention, the term "consisting of" is considered to be a preferred embodiment of the term "comprising" and excludes the presence of other elements. Hereinafter, when a group is defined to comprise at least a certain number of embodiments, this should also be understood to disclose a group that preferably consists of only these embodiments.

For the purposes of the present invention, the term "obtained" is considered to be a preferred embodiment of the term "obtainable". Below, if, for example, a cell or organism is defined as obtainable by a particular method, this should be understood to also disclose the cell or organism obtained by this method.

When an indefinite or definite article is used when referring to a singular noun such as "a", "an" or "the", this includes the plural of that noun unless specifically stated otherwise.

The term "pharmaceutical composition" as used herein refers to any composition containing a chemical or biological substance or active ingredient, which is intended for use in the medical cure, treatment, or prevention of disease, and is in a form such that the active ingredient is effective. In particular, a pharmaceutical composition does not contain excipients that are unacceptably toxic to the subject to which the composition is administered. A pharmaceutical composition is sterile, i.e., aseptic, and free of all living microorganisms and their spores. The pharmaceutical compositions used in the present invention are liquid and stable.

In the "liquid composition", the pharma- ceutically active agent, e.g., an anti-PD1 antibody, can be combined with various excipients to ensure a stable active drug after storage. In one embodiment, the liquid pharmaceutical composition used in the present invention is not lyophilized, i.e., the manufacturing method does not include a lyophilization step and the composition is not lyophilized for storage. The liquid composition can be stored in vials, IV bags, ampoules, cartridges, and prefilled or ready-to-use syringes.

In another embodiment, the liquid composition is lyophilized after its preparation. The terms "lyophilization," "lyophilized," and "freeze-dried" refer to a process in which the material to be dried is first frozen and then the ice or freezing solvent is removed by sublimation in a vacuum environment. Preferably, the lyophilized formulation is prepared by freeze-drying the liquid pharmaceutical composition of the present invention. Those skilled in the art are aware of lyophilization protocols.

A "stable" liquid composition is one in which the anti-PD1 antibody contained therein essentially retains its physical and/or chemical stability and/or biological activity upon storage for a particular period of time. Preferably, the composition essentially retains its physical and chemical stability, as well as its biological activity upon storage. Various analytical techniques for measuring protein stability are available in the art and are reviewed, for example, in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed, Marcel Dekker, Inc, New York, New York, Pubs (1991) and Jones, Adv Drug Delivery Rev, 1993, 10:29-90. For example, stability can be measured at a selected temperature over a selected period of time. Stability can be assessed qualitatively and/or quantitatively in a variety of different ways, including assessing aggregate formation (e.g., using size exclusion chromatography, by measuring turbidity, and/or by visual inspection), assessing charge heterogeneity using cation exchange chromatography or capillary zone electrophoresis, amino- or carboxy-terminal sequence analysis, mass spectrometry, SDS-PAGE analysis to detect aggregated or fragmented molecules, peptide map (e.g., trypsin or LYS-C) analysis, assessing antagonist biological activity or binding, etc.

Preferably, the pharmaceutical composition is stable for at least 1-2 weeks at a temperature of about 40° C., and/or for at least 3 months, preferably 6 months or 9 months, more preferably 1 year at a temperature of about 5° C., and/or for at least 2 weeks or 1 month at a temperature of about 25° C. Furthermore, the formulation is preferably stable after freezing the formulation (e.g., to −80° C.) and thawing at 25° C., e.g., after 1, 2, 3 or 4 cycles of freezing and thawing, as described in the Examples herein.

For example, in the pharmaceutical composition of the present invention, the percentage of high molecular weight species of anti-PD1 antibody relative to the total amount of anti-PD1 antibody as measured by size exclusion chromatography is 10% or less, preferably 5% or less, more preferably 3% or less, and most preferably 2% or less after storage for 2 weeks at a temperature of about 40°C.

Alternatively, in the pharmaceutical composition of the present invention, the percentage of high molecular weight species of anti-PD1 antibody relative to the total amount of anti-PD1 antibody as measured by size exclusion chromatography is 10% or less, preferably 5% or less, more preferably 3% or less, and most preferably 2% or less after storage at a temperature of about 5°C for one month.

Alternatively, in the pharmaceutical composition of the present invention, the percentage of high molecular weight species of anti-PD1 antibody relative to the total amount of anti-PD1 antibody as measured by size exclusion chromatography is 10% or less, preferably 5% or less, more preferably 3% or less, and most preferably 2% or less after storage at a temperature of about 5°C for 3 months, 6 months, or 9 months.

Furthermore, in the pharmaceutical composition of the present invention, the percentage of glycated species of anti-PD1 antibody relative to the total amount of anti-PD1 antibody measured by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) is 2.3% or less, preferably 2.0% or less, and more preferably 1.8% or less after storage at a temperature of about 40°C for one month.

A "buffer" is an aqueous solution consisting of a mixture of a weak acid and its conjugate base, or vice versa, that resists changes in its pH and keeps the pH at a nearly constant value. The buffers of the present invention preferably have a pH in the range of about 5.0 to about 5.8, preferably about 5.2 to about 5.7, more preferably about 5.4 to 5.7, and most preferably about 5.5.

The buffer used in the present invention is a histidine-containing buffer or a citrate buffer. In one embodiment, the citrate buffer is the only buffer present in the pharmaceutical composition of the present invention. Preferably, the histidine-containing buffer is the only buffer present in the liquid pharmaceutical composition of the present invention. In particular, the liquid pharmaceutical composition of the present invention does not include a mixture of a histidine buffer and a citrate buffer.

In one embodiment, the buffer is a citrate buffer. The citrate buffer is prepared by mixing citric acid with a citrate salt such as sodium citrate, or by mixing citric acid with a base such as sodium hydroxide, arginine and/or lysine. The citrate buffer has a concentration of 1 mM to 50 mM, preferably 2 mM to 40 mM, more preferably 3 mM to 30 mM, even more preferably 4 mM to 20 mM, and most preferably 5 mM to 10 mM. In one embodiment, the citrate buffer has a concentration of 5 mM. In one embodiment, the citrate buffer has a concentration of 10 mM. In one embodiment, the citrate buffer has a concentration of 2 mM.

According to the present invention, the citrate buffer has a pH in the range of about 5.0 to 5.8, preferably about 5.1 to 5.7, more preferably about 5.2 to 5.6 or about 5.3 to 5.6, and most preferably about 5.5.

In one embodiment, the citrate buffer comprises citric acid and sodium citrate at a concentration of 10 mM. In another embodiment, the citrate buffer comprises citric acid and sodium citrate at a concentration of 10 mM and a pH of 5.5.

In one embodiment, the citrate buffer has a concentration of 10 mM and a pH of 5.5.
In one embodiment, the citrate buffer comprises citric acid and citrate salt at a concentration of 10 mM. In another embodiment, the citrate buffer comprises citric acid and citrate salt at a concentration of 10 mM and a pH of 5.5. The pH can be adjusted to pH 5.5 by adding HCl/sodium hydroxide, arginine/arginine HCl, and/or lysine/lysine HCl.

In one embodiment, the citrate buffer comprises citric acid and sodium citrate at a concentration of 5 mM. In another embodiment, the citrate buffer comprises citric acid and sodium citrate at a concentration of 5 mM and a pH of 5.5.

In one embodiment, the citrate buffer comprises citric acid and citrate salt at a concentration of 5 mM. In another embodiment, the citrate buffer comprises citric acid and citrate salt at a concentration of 5 mM and a pH of 5.5. The pH can be adjusted to pH 5.5 by adding HCl/sodium hydroxide, arginine/arginine HCl, and/or lysine/lysine HCl.

The terms "histidine-containing buffer" and "histidine buffer" are used interchangeably herein and refer to a buffer that contains histidine. Examples of histidine buffers include histidine chloride, histidine hydrochloride, histidine acetate, histidine phosphate, and histidine sulfate. A preferred histidine buffer of the present invention further comprises L-histidine. Even more preferably, the histidine buffer of the present invention comprises histidine hydrochloride, and most preferably comprises histidine hydrochloride and L-histidine.

Preferably, the histidine buffer or histidine hydrochloride buffer or histidine hydrochloride/L-histidine buffer has a pH in the range of about 5.2 to 5.8, preferably about 5.3 to 5.7, more preferably 5.4 to 5.6, and most preferably a pH of about 5.5.

In a particularly preferred embodiment, the histidine-containing buffer contains histidine hydrochloride/L-histidine at a concentration of 5-30 mM, preferably 7-20 mM, more preferably 8-15 mM, and most preferably 10 mM.

In another particularly preferred embodiment, the buffer is histidine hydrochloride/L-histidine having a concentration of 10 mM and a pH of 5.5.
As used herein, "surfactant" refers to an amphipathic compound, i.e., a compound that contains both hydrophobic and hydrophilic groups that reduce the surface tension (or interfacial tension) between two liquids or between a liquid and a solid. A "non-ionic surfactant" does not have a charged group in its head. The formation of insoluble particles during freeze/thaw cycles of an antibody-containing composition can be significantly inhibited by the addition of surfactants. Examples of "nonionic surfactants" include, for example, polyoxyethylene glycol alkyl ethers, such as octaethylene glycol monododecyl ether, pentaethylene glycol monododecyl ether; polyoxypropylene glycol alkyl ethers; glucoside alkyl ethers, such as decyl glucoside, lauryl glucoside, octyl glucoside; polyoxyethylene glycol octylphenol ethers, such as Triton X-100; polyoxyethylene glycol alkylphenol ethers, such as nonoxynol-9; glycerol alkyl esters, such as glyceryl laurate; polyoxyethylene glycol sorbitan alkyl esters, such as polysorbates; sorbitan alkyl esters, such as spans; cocamide MEA, cocamide DEA, dodecyl dimethylamine oxide; block copolymers of polyethylene glycol and polypropylene glycol, such as poloxamers; and polyethoxylated tallow amine (POEA). The liquid pharmaceutical compositions of the present invention may contain one or more of these surfactants in combination. In a preferred embodiment, the pharmaceutical composition of the present invention contains only one non-ionic surfactant.

Preferred non-ionic surfactants for use in the pharmaceutical compositions of the present invention are polysorbates such as polysorbate 20, 40, 60 or 80, particularly polysorbate 20 (i.e. Tween 20) or polysorbate 80 (i.e. Tween 80).

The concentration of the nonionic surfactant is in the range of 0.005-0.06% (w/v) based on the total volume of the composition, preferably in the range of 0.008-0.05% (w/v), and most preferably in the range of 0.01-0.04% (w/v).

In a preferred embodiment, the non-ionic surfactant is polysorbate 20. In a preferred embodiment, the non-ionic surfactant is polysorbate 20 having a concentration in the range of 0.05-0.6 mg/ml, preferably in the range of 0.08-0.5 mg/ml, and most preferably in the range of 0.1-0.4 mg/ml. In a preferred embodiment, the non-ionic surfactant is polysorbate 20 having a concentration of 0.1 mg/ml. In a preferred embodiment, the non-ionic surfactant is polysorbate 20 having a concentration of 0.2 mg/ml.

In another preferred embodiment, the non-ionic surfactant is polysorbate 80 having a concentration in the range of 0.05-0.6 mg/ml, preferably in the range of 0.08-0.5 mg/ml, more preferably in the range of 0.1-0.4 mg/ml, and most preferably 0.2 mg/ml.

In a particularly preferred embodiment, the non-ionic surfactant is polysorbate 80 having a concentration of 0.2 mg/ml.
In one embodiment, the non-ionic surfactant is polysorbate 20 having a concentration of 0.4 mg/ml. In one embodiment, the non-ionic surfactant is polysorbate 20 having a concentration of 0.1 mg/ml.

In one embodiment, the pharmaceutical composition comprising an anti-human PD1 antibody, a citrate buffer, a non-ionic surfactant, and one or more amino acids other than histidine does not contain sugar. In one embodiment, the pharmaceutical composition comprising an anti-human PD1 antibody, a citrate buffer, a non-ionic surfactant, and one or more amino acids other than histidine does not contain sucrose. In one embodiment, the pharmaceutical composition comprising an anti-human PD1 antibody, a citrate buffer, a non-ionic surfactant, and one or more amino acids other than histidine does not contain trehalose.

The liquid pharmaceutical composition of the present invention comprises one or more amino acids other than histidine. Amino acids are organic compounds containing an amino group and a carboxyl group and a side chain unique to each amino acid. Amino acids that may be present in the liquid pharmaceutical composition of the present invention may be selected from the group consisting of arginine, lysine, aspartic acid, glutamic acid, serine, threonine, asparagine, glutamine, cysteine, glycine, proline, alanine, valine, isoleucine, methionine, phenylalanine, tyrosine and tryptophan. In one embodiment, the liquid pharmaceutical composition of the present invention comprises arginine. In one embodiment, the liquid pharmaceutical composition of the present invention comprises lysine. In one embodiment, the liquid pharmaceutical composition of the present invention comprises arginine and lysine. In one embodiment, arginine is the only amino acid present in the liquid pharmaceutical composition of the present invention. In one embodiment, lysine is the only amino acid present in the liquid pharmaceutical composition of the present invention. In one embodiment, the only amino acids present in the liquid pharmaceutical composition of the present invention are arginine and lysine.

Arginine and/or lysine may be added to the liquid pharmaceutical composition as the free base or its hydrochloride salt. In one embodiment, arginine is added as a mixture of free base arginine and the hydrochloride salt of arginine, i.e., arginine hydrochloride (arginine-HCl). In one embodiment, lysine is added as a mixture of free base lysine and the hydrochloride salt of lysine, i.e., lysine hydrochloride (lysine-HCl). In one embodiment, arginine is added as a mixture of free base arginine and the hydrochloride salt of arginine, i.e., arginine hydrochloride, and lysine is added as a mixture of free base lysine and the hydrochloride salt of lysine, i.e., lysine hydrochloride.

In one embodiment, arginine is added as a mixture of free base arginine and the hydrochloride salt of arginine, i.e., arginine hydrochloride (arginine-HCl), with the ratio of free base arginine to arginine-HCl being 1:5 to 1:12, preferably 1:7 to 1:12, more preferably 1:8 to 1:12, even more preferably 1:9 to 1:11, and most preferably 1:10.5.

In one embodiment, the liquid pharmaceutical composition of the present invention comprises arginine at a concentration of 100 mM to 350 mM, preferably 120 mM to 330 mM, more preferably 130 mM to 320 mM, and most preferably 150 mM to 300 mM. In one embodiment, the liquid pharmaceutical composition of the present invention comprises arginine at a concentration of 250 mM to 350 mM, preferably 260 mM to 340 mM, more preferably 270 mM to 330 mM, even more preferably 280 mM to 320 mM, and most preferably 290 mM to 310 mM. In one embodiment, the liquid pharmaceutical composition of the present invention comprises arginine at a concentration of 150 mM. In one embodiment, the liquid pharmaceutical composition of the present invention comprises arginine at a concentration of 250 mM. In one embodiment, the liquid pharmaceutical composition of the present invention comprises arginine at a concentration of 300 mM. In one embodiment, the liquid pharmaceutical composition of the present invention contains 26 mM L-arginine and 274 mM L-arginine-HCl.

In one embodiment, the liquid pharmaceutical composition of the present invention comprises lysine at a concentration of 100 mM to 300 mM, preferably 120 mM to 280 mM, more preferably 130 mM to 270 mM, and most preferably 150 mM to 250 mM. In one embodiment, the liquid pharmaceutical composition of the present invention comprises lysine at a concentration of 150 mM. In one embodiment, the liquid pharmaceutical composition of the present invention comprises lysine at a concentration of 250 mM.

In one embodiment, the liquid pharmaceutical composition of the present invention comprises arginine and lysine at a total concentration of 100 mM to 350 mM, preferably 150 mM to 330 mM, more preferably 180 mM to 310 mM, and most preferably 300 mM.

In one embodiment, the liquid pharmaceutical composition of the present invention comprises 100 mM to 200 mM arginine and 100 mM to 200 mM lysine. In one embodiment, the liquid pharmaceutical composition of the present invention comprises 120 mM to 180 mM arginine and 120 mM to 180 mM lysine. In one embodiment, the liquid pharmaceutical composition of the present invention comprises 130 mM to 170 mM arginine and 130 mM to 170 mM lysine. In one embodiment, the liquid pharmaceutical composition of the present invention comprises 140 mM to 160 mM arginine and 140 mM to 160 mM lysine. In one embodiment, the liquid pharmaceutical composition of the present invention comprises 150 mM arginine and 150 mM lysine.

In one embodiment, the liquid pharmaceutical composition of the present invention comprises a sugar alcohol. A sugar alcohol is an organic compound derived from a sugar that contains a hydroxy group attached to each carbon atom. Suitable sugar alcohols include glycerol, mannitol, sorbitol, and xylitol.

Preferably, the sugar alcohol is glycerol. The concentration of glycerol in the liquid pharmaceutical composition of the present invention is 50 mM to 200 mM, preferably the concentration of glycerol is 60 mM to 180 mM, more preferably the concentration of mannitol or sorbitol is 70 mM to 150 mM or 80 mM to 120 mM, and most preferably the concentration of glycerol is 100 mM. In one embodiment, the liquid pharmaceutical composition of the present invention comprises 100 mM glycerol.

In one embodiment, the liquid pharmaceutical composition of the present invention does not contain mannitol.In one embodiment, the liquid pharmaceutical composition of the present invention does not contain sorbitol.
In one embodiment, the liquid pharmaceutical composition of the present invention does not include sodium chloride. In one embodiment, the liquid pharmaceutical composition of the present invention does not include any sodium salt. In one embodiment, the liquid pharmaceutical composition of the present invention does not include any inorganic salt. As used herein, "inorganic salt" refers to an ionic compound that has osmoregulatory properties. An inorganic salt, such as sodium chloride (NaCl), can dissociate into its constituent ions in solution, i.e., NaCl dissociates into Na+ and Cl- ions, both of which affect the osmotic pressure, i.e., osmolality, of a solution. Exemplary inorganic salts that are not present in the liquid pharmaceutical composition of the present invention are potassium chloride, calcium chloride, sodium chloride, sodium phosphate, potassium phosphate, and sodium bicarbonate.

In one embodiment, the liquid pharmaceutical composition of the present invention does not contain EDTA. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain pentetic acid. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain EDTA and pentetic acid. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain any chelating agent. A chelating agent is capable of forming at least one bond with a metal atom. Chelating agents are typically multidentate ligands, which may be used in the composition as stabilizers to complex with species that may otherwise promote instability. Exemplary chelating agents include aminopolycarboxylic acids, hydroxyaminocarboxylic acids, N-substituted glycines, 2-(2-amino-2-oxocthyl)aminoethanesulfonic acid (BES), deferoxamine (DEF), niacinamide, desoxycholate, ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), nitrilotriacetic acid (NTA), N-2-acetamido-2-iminodiacetic acid (ADA), bis(aminoethyl) glycol ether, N,N,N',N'-tetraacetic acid (EGTA). , trans-diaminocyclohexanetetraacetic acid (DCTA), N-hydroxyethyliminodiacetic acid (HIMDA), N,N-bis-hydroxyethylglycine (bicine), N-(trishydroxymethylmethyl)glycine (tricine), glycylglycine, sodium desoxycholate, ethylenediamine; propylenediamine; diethylenetriamine; triethylenetetraamine (triene), ethylenediaminetetraacetoEDTA; disodium EDTA, EDTA, calcium EDTA, oxalic acid, and malate. Histidine and citrate are not considered chelating agents in the present invention.

In one embodiment, the liquid pharmaceutical composition of the present invention does not contain methionine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain L-methionine or L-methionine-HCl. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain any antioxidants. Antioxidants are compounds that inhibit oxidation by reacting with oxidizing agents. Histidine is not considered an antioxidant in the present invention.

In one embodiment, the liquid pharmaceutical composition of the present invention does not contain proline. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain glycine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain glutamic acid. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain serine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain tyrosine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain tryptophan. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain leucine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain phenylalanine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain threonine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain aspartate. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain asparagine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain glutamine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain alanine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain cysteine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain isoleucine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain valine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain any amino acid in addition to arginine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain any amino acid in addition to lysine. In one embodiment, the liquid pharmaceutical composition of the present invention does not contain any amino acids in addition to arginine and lysine.

In one embodiment, the liquid pharmaceutical composition of the present invention is free of EDTA and proline.In one embodiment, the liquid pharmaceutical composition of the present invention is free of DTPA and methionine.
The term "sugar" refers to an organic compound that contains only carbon, hydrogen, and oxygen, usually with an atomic ratio of hydrogen:oxygen of 2:1, and has the empirical formula Cm(HO)n. The term "sugar" includes monosaccharides, disaccharides, oligosaccharides, and polysaccharides. Examples of sugars include glucose, fructose, galactose, xylose, ribose, sucrose, mannose, lactose, maltose, trehalose, starch, and glycogen. Preferably, the sugar is a non-reducing sugar. A non-reducing sugar is a sugar that does not contain a free aldehyde or ketone group and therefore cannot act as a reducing agent. Preferably, the non-reducing sugar is selected from sucrose and trehalose.

In one embodiment, the sugar is trehalose. Trehalose is a non-reducing sugar. It is a disaccharide formed by a 1,1-glycosidic bond between a glucose unit and a fructose unit. Preferably, the dihydrate form of trehalose is used. The concentration of trehalose dihydrate in the liquid pharmaceutical composition of the present invention is 100 mM to 300 mM, preferably the concentration of trehalose dihydrate is 120 mM to 280 mM, more preferably the concentration of trehalose dihydrate is 150 mM to 250 mM or 150 mM to 205 mM, and most preferably the concentration of trehalose dihydrate is 150 mM or 205 mM.

In one embodiment, the sugar is sucrose. Sucrose is a non-reducing sugar. It is a disaccharide formed by a 1,2-glycosidic bond between two α-glucose units. The concentration of sucrose in the liquid pharmaceutical composition of the present invention is 100 mM to 300 mM, preferably the concentration of sucrose is 120 mM to 280 mM, more preferably the concentration of sucrose is 150 mM to 250 mM, and most preferably the concentration of sucrose is 205 mM. A sucrose concentration of 205 mM is equivalent to 70 mg/ml sucrose.

The term "antibody" or "immunoglobulin" is used herein in the broadest sense and includes full length antibodies, genetically engineered antibodies, recombinant antibodies, multivalent antibodies, monoclonal antibodies, polyclonal antibodies, bispecific antibodies, multispecific antibodies, chimeric antibodies, humanized antibodies, fully human antibodies, and fragments of such antibodies so long as they remain functional and exhibit the desired biological activity. The "biological activity" of an antibody refers to the ability of the antibody to bind an antigen and result in a biological response that can be measured in vitro or in vivo.

A full length antibody comprises an antigen-binding variable region of a light chain (VL) and a heavy chain (VH), a light chain constant region (CL) and heavy chain constant domains CH1, CH2 and CH3.
The term "antibody fragment" or "antigen-binding fragment" is used herein in the broadest sense and includes a portion of a full-length antibody, preferably including the antigen-binding or variable region thereof. Antibody fragments retain the original specificity of the parent immunoglobulin. Examples of antibody fragments include, for example, Fab, Fab', F(ab')2, and Fv fragments, diabodies, linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragment(s).

A "monoclonal antibody" is an antibody specific for a single epitope of an antigen, i.e., an antibody against a single determinant on the antigen. Methods for producing monoclonal antibodies are known to those skilled in the art.

The term "recombinant antibody" refers to any antibody prepared, expressed, produced or isolated by recombinant means, e.g., an antibody isolated from a transgenic host cell, e.g., an NS0 or CHO cell, or from an animal transgenic for an immunoglobulin gene, or an antibody expressed using a recombinant expression vector transfected into a host cell, e.g., an SP2/0 mouse myeloma cell.

"Humanized antibodies" are human antibodies whose antigen-binding portions (CDRs) are derived from a non-human species, such as mouse, and have different specificity compared to the parent immunoglobulin. The CDR protein sequences can be modified to increase their similarity to antibody variants naturally produced in humans.

A "fully human antibody" is one in which all portions of the antibody, including the antigen-binding portions (CDRs), are derived from humans.
The term "anti-PD1 antibody" refers to an antibody that specifically binds to cell death protein 1 (PD-1) and inhibits binding of PD-1 to its ligand PD-L1, and optionally inhibits binding of PD-1 to its ligands PD-L1 and PD-L2. The anti-PD1 antibody thereby negates the inhibitory effect of PD-1/PD-L1 interaction on T cells. Known anti-PD1 antibodies include, but are not limited to, pembrolizumab, nivolumab, cemiplimab, and cetrelimab.

Pembrolizumab (also known as MK-3475, SCH900475 and lambrolizumab) is a humanized IgG4 mAb having the structure described in WHO Drug Information, Vol. 27, No. 2, pp. 161-162 (2013) and contains the heavy and light chain amino acid sequences and CDRs described in Table 2 of WO 2018/204368. Pembrolizumab is approved, among others, for the treatment of patients with unresectable or metastatic melanoma, and for the treatment of patients with recurrent or metastatic head and neck squamous cell carcinoma (HNSCC), classical Hodgkin lymphoma (cHL), urothelial carcinoma, gastric cancer, microsatellite instability-high (MSI-H) cancer and non-small cell lung cancer. The current commercially available pembrolizumab formulation contains 10 mM histidine, 70 mg/ml sucrose, 0.2 mg/ml polysorbate 80, and water for injection (pH of the formulation is 5.5) and is supplied at a concentration of 25 mg/ml.

Nivolumab (also known as ONO-4538, BMS-936558, MDX1106) is a fully human monoclonal IgG4 antibody containing the heavy and light chain amino acid sequences and CDRs set out in Table 2 of WO 2018/204368. Pembrolizumab is approved for the treatment of patients with melanoma, renal carcinoma, non-small cell lung cancer and urothelial carcinoma. The current marketed formulation of nivolumab contains 30 mg/ml mannitol, 0.008 mg/ml pentetic acid, 0.2 mg/ml polysorbate 80, 2.92 mg/ml sodium chloride, 5.88 mg/ml sodium citrate dihydrate, and water for injection (pH of the formulation 6.0) and is supplied at a concentration of 10 mg/ml.

In one embodiment, the liquid pharmaceutical composition does not comprise an anti-LAG3 antibody. In one embodiment, the antibody is not a bispecific antibody.
In one embodiment, the anti-human PD-1 antibody is the only antibody present in the liquid pharmaceutical composition. In one embodiment, pembrolizumab is the only antibody present in the liquid pharmaceutical composition.

In one embodiment, the anti-human PD-1 antibody is the only pharma- ceutical active agent present in the liquid pharmaceutical composition. In one embodiment, pembrolizumab is the only pharma-ceutical active agent present in the liquid pharmaceutical composition.

The concentration of the anti-PD1 antibody in the pharmaceutical composition of the present invention is typically 10 to 80 mg/ml, preferably 15 to 70 mg/ml or 15 to 60 mg/ml, more preferably 20 to 50 mg/ml or 20 to 40 mg/ml, and most preferably 25 mg/ml.

The pharmaceutical composition of the present invention can be used for the treatment of cancer, in particular for the treatment of melanoma, non-small cell lung cancer, classical Hodgkin's lymphoma, urothelial carcinoma, head and neck squamous cell carcinoma, renal cell carcinoma, colorectal cancer, esophageal cancer, small cell lung cancer, high-frequency microsatellite instability or mismatch repair deficient cancer, primary mediastinal large B-cell lymphoma, gastric or gastroesophageal junction adenocarcinoma, hepatocellular carcinoma, Merkel cell carcinoma, endometrial cancer, oncogene hypermutation cancer, cutaneous squamous cell carcinoma, triple-negative breast cancer or cervical cancer.

The pharmaceutical compositions of the present invention may contain further active agents, particularly further antitumor agents such as chemotherapeutic agents. Examples of such chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide; alkylsulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa and uredopa; ethylenimines and methylamelanamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolomelamine; acetogenins (particularly bullatacin and bullatacinone); camptothecin (including the synthetic analogue topotecan); bryostatin; kallistatin; CC-1065 (including its synthetic analogues adozelesin, carzelesin and bizelesin); cryptophycins (particularly cryptophycins, phycin 1 and cryptophycin 8); dolastatins; duocarmycins (including synthetic analogs, KW-2189 and CBI-TMI); eleutherobin; pancratistatin; sarcodictyin; spongistatins; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembitine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; enediyne antibiotics (e.g. calicheamicin, especially calicheamicin gamma ll); gammall), calicheamicin fil; dynemicins including dynemicin A; bisphosphonates such as clodronate; esperamicin; and neocarzinostatin chromophores and related chromoprotein enediyne antibiotics chromomophores), aclacinomycins, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, carabicin, ca Minomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcelomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycin , peplomycin, potfilomycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; fludarabine, 6-mercaptopurine, thiamiprine, Purine analogues such as thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; florinic acid folic acid replenishers such as aceglatone, aldophosphamide glycosides, aminolevulinic acid, eniluracil, amsacrine, bestravcil, bisantrene, edatraxate, defofamine, demecolcine, diaziquone, elformitin, elliptinium acetate, epothilone, etoglucide, gallium nitrate, hydroxyurea, lentinan, lonidamine, maytansinoids such as maytansine and ansamitocins inoids; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; razoxane; rhizoxin; schizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2''-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A) A), Roridin A and anguidine; urethane; vindesine; dacarbazine; mannommustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, such as paclitaxel and doxetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine ; methotrexate; platinum analogues such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; the topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Also included are anti-hormonal agents that act to regulate or inhibit hormone action on tumors, such as anti-estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen, raloxifene, droloxifene, 4-hydroxytamoxifen, trioxyphene, keoxifene, LY117018, onapristone, and toremifene (Fareston); aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazole, aminoglutethimide, megestrol acetate, exemestane, formestane, fadrozole, vorozole, letrozole, and anastrozole; and anti-androgens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharma- ceutically acceptable salts, acids, or derivatives of any of the above.

In an alternative embodiment, the pharmaceutical composition of the present invention may be administered in combination with any of the chemotherapeutic agents listed above, but the chemotherapeutic agent is present in a separate pharmaceutical composition.

In an alternative embodiment, the pharmaceutical composition of the present invention is administered with pemetrexed and platinum-based chemotherapy. In an alternative embodiment, the pharmaceutical composition of the present invention is administered with pemetrexed and platinum-based chemotherapy in the treatment of non-small cell lung cancer. In an alternative embodiment, the pharmaceutical composition of the present invention is administered with carboplatin and either paclitaxel or nab-paclitaxel. In an alternative embodiment, the pharmaceutical composition of the present invention is administered with carboplatin and either paclitaxel or nab-paclitaxel in the treatment of non-small cell lung cancer.

In an alternative embodiment, the pharmaceutical composition of the present invention is administered with platinum-based chemotherapy and 5-fluorouracil. In an alternative embodiment, the pharmaceutical composition of the present invention is administered with platinum-based chemotherapy and 5-fluorouracil in the treatment of head and neck squamous cell carcinoma.

In an alternative embodiment, the pharmaceutical composition of the present invention is administered with axitinib. In an alternative embodiment, the pharmaceutical composition of the present invention is administered with axitinib in the treatment of renal cell carcinoma.

In an alternative embodiment, the pharmaceutical composition of the present invention is administered with platinum-based chemotherapy and fluoropyrimidine-based chemotherapy. In an alternative embodiment, the pharmaceutical composition of the present invention is administered with platinum and fluoropyrimidine-based chemotherapy in the treatment of esophageal cancer.

In an alternative embodiment, a pharmaceutical composition of the present invention is administered with lenvatinib. In an alternative embodiment, a pharmaceutical composition of the present invention is administered with lenvatinib in the treatment of endometrial cancer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM to 350 mM arginine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml of anti-human PD1 antibody; 250 mM to 350 mM arginine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml of anti-human PD1 antibody; 280 mM to 320 mM arginine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM to 300 mM lysine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM to 350 mM of arginine and lysine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml of anti-human PD1 antibody; 100 mM to 200 mM arginine; 100 mM to 200 mM lysine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml anti-human PD1 antibody; 100 mM to 350 mM arginine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml of anti-human PD1 antibody; 250 mM to 350 mM arginine; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml anti-human PD1 antibody; 280 mM to 320 mM arginine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml anti-human PD1 antibody; 100 mM to 300 mM lysine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml anti-human PD1 antibody; 100 mM to 350 mM arginine and lysine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml anti-human PD1 antibody; 100 mM to 200 mM arginine; 100 mM to 200 mM lysine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 100 mM to 350 mM arginine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 250 mM to 350 mM arginine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 280 mM to 320 mM arginine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 100 mM to 300 mM lysine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 100 mM to 350 mM arginine and lysine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 100 mM to 200 mM arginine; 100 mM to 200 mM lysine; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 100 mM to 350 mM arginine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 250 mM to 350 mM arginine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 280 mM to 320 mM arginine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 100 mM to 300 mM lysine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 100 mM to 350 mM arginine and lysine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml pembrolizumab; 100 mM to 200 mM arginine; 100 mM to 200 mM lysine; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 300 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 300 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 150 mM arginine; 150 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 300 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 300 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 150 mM arginine; 150 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 300 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 300 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 150 mM arginine; 150 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 300 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 300 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 150 mM arginine; 150 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 300 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 300 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 150 mM arginine; 150 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 300 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 300 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 150 mM arginine; 150 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 300 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 300 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 150 mM arginine; 150 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant; and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 300 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 300 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 150 mM-200 mM arginine; 150 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.2 mg/ml of polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml anti-human PD1 antibody; 250 mM-350 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml anti-human PD1 antibody; 280 mM-320 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml anti-human PD1 antibody; 100 mM-300 mM lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml anti-human PD1 antibody; 100 mM-350 mM arginine and lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.2 mg/ml of polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml anti-human PD1 antibody; 250 mM-350 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml anti-human PD1 antibody; 280 mM-320 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml anti-human PD1 antibody; 100 mM-300 mM lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.2 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM of arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml of anti-human PD1 antibody; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 10-80 mg/ml pembrolizumab; 250 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 10-80 mg/ml pembrolizumab; 280 mM-320 mM arginine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 10-80 mg/ml pembrolizumab; 100 mM-300 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-350 mM arginine and lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 10-80 mg/ml pembrolizumab; 100 mM-200 mM arginine; 100 mM-200 mM lysine; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml anti-PD1 antibody; 300 mM arginine; 0.2 mg/ml polysorbate 80 and 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 25 mg/ml anti-PD1 antibody; 300 mM arginine; 0.2 mg/ml polysorbate 80 and 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml pembrolizumab; 300 mM arginine; 0.2 mg/ml polysorbate 80 and 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml pembrolizumab; 300 mM arginine; 0.2 mg/ml polysorbate 80 and 5 or 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml anti-PD1 antibody; 250 mM lysine; 0.2 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml anti-PD1 antibody; 250 mM lysine; 0.2 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml pembrolizumab; 250 mM lysine; 0.2 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.0-5.8).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml pembrolizumab; 250 mM lysine; 0.2 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 25 mg/ml anti-PD1 antibody; 150 mM arginine; 150 mM lysine; 0.2 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml anti-PD1 antibody; 150 mM arginine; 150 mM lysine; 0.2 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml pembrolizumab; 150 mM arginine; 150 mM lysine; 0.2 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.0-5.8).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml pembrolizumab; 150 mM arginine; 150 mM lysine; 0.2 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml pembrolizumab; 300 mM arginine/arginine HCl; 0.2 mg/ml polysorbate 80 and 10 mM citric acid monohydrate, pH 5.5.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 25 mg/ml pembrolizumab; 26 mM L-arginine; 274 mM L-arginine HCl; 0.2 mg/ml polysorbate 80 and 10 mM citric acid monohydrate, pH 5.5.

The pharmaceutical composition can be supplied in a vial or a pre-filled syringe. The pharmaceutical composition can be administered by intravenous infusion, for example over a period of up to 30 minutes.
Alternatively, the pharmaceutical composition may be administered by subcutaneous injection. In this case, the concentration of the anti-PD1 antibody, preferably pembrolizumab, is between 80 mg/ml and 180 mg/ml, more preferably between 90 mg/ml and 170 mg/ml, most preferably between 100 mg/ml and 165 mg/ml. In one embodiment, the concentration of the anti-PD1 antibody, preferably pembrolizumab, is 100 mg/ml, 150 mg/ml or 165 mg/ml.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 90 mg/ml to 350 mg/ml, preferably 100 mg/ml to 165 mg/ml, of an anti-human PD-1 antibody, preferably pembrolizumab; trehalose dihydrate; a non-ionic surfactant and a citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 90 mg/ml to 350 mg/ml, preferably 100 mg/ml to 165 mg/ml, of an anti-human PD-1 antibody, preferably pembrolizumab; trehalose dihydrate; polysorbate 80 and a citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 90 mg/ml to 350 mg/ml, preferably 100 mg/ml to 165 mg/ml, of an anti-human PD-1 antibody, preferably pembrolizumab; trehalose dihydrate; a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 90 mg/ml to 350 mg/ml, preferably 100 mg/ml to 165 mg/ml, of an anti-human PD-1 antibody, preferably pembrolizumab; trehalose dihydrate; polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the liquid pharmaceutical composition comprising 90 mg/ml to 350 mg/ml, preferably 100 mg/ml to 165 mg/ml of an anti-human PD-1 antibody, preferably pembrolizumab, may further contain arginine. When arginine is present, it may be present at a concentration of 10 mM to 200 mM. When arginine is present in the liquid pharmaceutical composition comprising 90 mg/ml to 350 mg/ml, preferably 100 mg/ml to 165 mg/ml of an anti-human PD-1 antibody, preferably pembrolizumab, the concentration of trehalose may be reduced to adjust osmolality.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 90 mg/ml to 350 mg/ml, preferably 100 mg/ml to 165 mg/ml, of an anti-human PD-1 antibody, preferably pembrolizumab; arginine; polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml to 180 mg/ml of an anti-human PD1 antibody; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of a citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80, and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of an anti-human PD1 antibody; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of a citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of an anti-human PD1 antibody; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of a citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80, and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml to 180 mg/ml of pembrolizumab; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80 mg/ml-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of pembrolizumab; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of pembrolizumab; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of pembrolizumab; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml pembrolizumab; 150 mM to 250 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml pembrolizumab; 150 mM to 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80 mg/ml to 180 mg/ml of pembrolizumab; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80 and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80 and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80 and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml of an anti-human PD1 antibody; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of a citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM to 250 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80 and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM to 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80 and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80 and 1 to 50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM to 250 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM to 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM to 250 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM to 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM to 250 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM to 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM to 250 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM to 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100, 150 or 165 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml polysorbate 80 and 1 to 50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.2 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.2 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.2 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.2 mg/ml of a non-ionic surfactant and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.2 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.2 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.2 mg/ml of polysorbate 80 and 1-50 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.2 mg/ml non-ionic surfactant and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 1-50 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 5 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 10 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 10 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 2 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 2 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 5 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 5 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 10 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 10 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant, and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant, and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 2 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 2 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 5 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 5 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 5 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-250 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM-205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80, and 10 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml of anti-human PD1 antibody; 150 mM or 205 mM of trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of polysorbate 80 and 10 mM of citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 100 mM-350 mM arginine; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml to 0.4 mg/ml of a non-ionic surfactant and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 2 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 2 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 5 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 80-180 mg/ml pembrolizumab; 150 mM-250 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 80-180 mg/ml pembrolizumab; 150 mM-205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 80-180 mg/ml pembrolizumab; 150 mM or 205 mM trehalose dihydrate; 0.1 mg/ml-0.4 mg/ml polysorbate 80 and 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100 mg/ml, 150 mg/ml or 165 mg/ml of anti-PD1 antibody; 205 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 3, 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100 mg/ml, 150 mg/ml or 165 mg/ml of anti-PD1 antibody; 205 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 3, 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100 mg/ml, 150 mg/ml or 165 mg/ml pembrolizumab; 205 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 100 mg/ml, 150 mg/ml or 165 mg/ml pembrolizumab; 205 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 5 or 10 mM citrate buffer (pH 5.5).

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100 mg/ml, 150 mg/ml or 165 mg/ml of anti-PD1 antibody; 150 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 3, 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.5) comprising 100 mg/ml, 150 mg/ml or 165 mg/ml of anti-PD1 antibody; 150 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 3, 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition (pH 5.0-5.8) comprising 100 mg/ml, 150 mg/ml or 165 mg/ml pembrolizumab; 150 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 5 or 10 mM citrate buffer.

In one embodiment, the present invention relates to a liquid pharmaceutical composition comprising 100 mg/ml, 150 mg/ml or 165 mg/ml pembrolizumab; 150 mM trehalose dihydrate; 0.2 mg/ml polysorbate 80 and 5 or 10 mM citrate buffer (pH 5.5).

While the invention has been illustrated and described in detail in the drawings and the foregoing description, such illustration and description are to be considered as specific or exemplary and not restrictive. The invention is not limited to the disclosed embodiments. Other variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing the claimed invention, from a study of the drawings, the disclosure, and the dependent claims.

The detailed description is merely illustrative in nature and is not intended to limit the application and use. The following examples further illustrate the present invention but are not intended to limit the scope of the present invention. Various changes and modifications can be made by those skilled in the art based on the description of the present invention, and such changes and modifications are also included in the present invention.

[Example]
I. Formulation Development of 25 mg/mL Pembrolizumab Formulation Example 1: Screening of Pembrolizumab Formulations 1. Sample Preparation Two independent Design of Experiments (DoE) studies were conducted to screen for suitable pembrolizumab formulations.

Initially, DoE studies (including formulation numbers (1) to (32) in Table 1) were prepared and evaluated in a D-optimal design to obtain the first-order and second-order effects of the components as well as selected interactions. As single components, histidine (covering a concentration range of 0-30 mM), citrate (0-30 mM), trehalose dihydrate (0-300 mM), mannitol (0-300 mM), polysorbate 20 (0-0.2 mg/mL) and pH (5.0-6.5) were evaluated for their stabilizing effects. The center point (number (7)) was included in the study as a triplicate preparation.

A second DoE study (including formulation nos. (33)-(55) in Table 1) was prepared and also evaluated in D-optimal design for first and second order effects of components and selected interactions. As single components, lysine HCl (covering a concentration range of 0-150 mM), arginine HCl (0-150 mM), proline (0-100 mM), glycine (0-75 mM) and glycerol (0-200 mM) were evaluated for their stabilizing effect. Core component buffer (10 mM citrate, pH 5.5) and 0.2 mg/ml polysorbate 20 were kept constant in all formulations. The center point (no. (39)) was included in the study as a duplicate preparation.

Using 25 mg/mL pembrolizumab as the starting material, 57 formulations listed in Table 1 were prepared by three-step dialysis. The dialyzed pembrolizumab was adjusted to 25 mg/mL ± 20% and sterile filtered at 0.22 μm. The formulations shown in Table 1 were first analyzed for protein concentration by UV-VIS spectroscopy at 280 nm and precise pH adjustment was performed before starting the accelerated aging stability program. Storage conditions are shown in Table 2. Briefly, samples were stored at 40°C/75% relative humidity (RH) for up to 2 weeks. Additionally, samples were subjected to two conditions of mechanical stress (overhead rotation, orbital shaking) as well as five freeze/thaw cycles (-80°C/+25°C). Pembrolizumab in Keytruda®-labeled formulation buffer was included in the study as a control sample (57) and was directly dialyzed with the other formulations shown in Table 1.

Table 1: Detailed information about the formulations prepared in this study (Example 1)

Table 2: Accelerated degradation of pembrolizumab formulations

Samples were analyzed by size-exclusion chromatography (SE-HPLC) for the presence of high molecular weight species (HMWS). Additionally, ion-exchange chromatography (IEX-HPLC) was used to detect modifications that result in charge heterogeneity.

2. Analysis of Protein Content by UV-VIS and Determination of pH and Osmolality All formulations prepared according to Table 1 were analyzed for their protein content as well as pH and osmolality before starting the stability study (t0). All samples met the acceptance criteria for protein content (25 mg/mL ± 20% pembrolizumab) and pH (target ± 0.2). No visible particles or color changes were detected in all samples.

3. Analysis of high molecular weight species (HMWS) by SE-HPLC.
Accelerated degradation study samples were diluted to a final concentration of 1 mg/mL with 20 mM histidine pH 5.5 and 5 μL was injected onto a TSKgel UP-SW3000 (Tosoh, 4.6×150 mm, 2 μm) column to detect high molecular weight species (HMWS) of pembrolizumab.

Proteins were eluted by isocratic elution using 0.1 M sodium phosphate buffer (pH 6.7) containing 0.05% (v/v) sodium azide at a flow rate of 0.35 mL/min at 25°C. Eluting species were detected at a wavelength of 280 nm and displayed on a graph showing the concentration of eluting species versus time. The elution profile showed a monomer peak with non-aggregated protein and multiple peaks of protein representing high molecular weight species (HMWS) of protein. The areas of all peaks were determined.

Table 3 summarizes the percentage of HMWS peak area relative to the total peak area of the eluting species for the formulations shown in Table 1. Each sample was measured in triplicate.
Table 3: Summary of HMWS determined using SE-HPLC

After 2 weeks of storage at 40°C, HMWS levels increased slightly to 0.8%-1.4% compared to t0 (0.7%) for all formulations tested in the design of the experimental set-up. For formulations (1)-(32), the effect of the buffer can be summarized as follows: Histidine is optimal at a molar concentration of 20-25 mM and a pH of 5.2-5.8.

For citrate, the optimum pH shifted slightly to lower pH values (approximately pH 5.0-5.6), and for high concentrations of citrate, a slight destabilizing effect was observed at high temperatures, which decreased with decreasing citrate molarity. Thus, histidine as well as citrate (especially when used at low concentrations up to 10 mM citrate) are the optimal buffer system for pembrolizumab.

For formulations (33)-(56), citrate buffer was used as the core component.
The sugar and sugar alcohol concentrations had only a small effect on the HMWS levels in formulations (1)-(32), as the formulations were robust over the range of sugar concentrations tested. Comparing formulations (33)-(56) containing amino acids and/or glycerol as stabilizing excipients, optimal conditions for pembrolizumab formulation were identified in samples (33), (36), (41), (42) and (45). Further evaluation revealed that a combination of high concentrations of arginine and/or lysine with 100 mM glycerol could achieve results comparable to the sugars and/or sugar alcohols used in formulations (1)-(32) in terms of HMWS levels. Formulation (56) containing poloxamer 188 provided results comparable to polysorbate 20 (PS20)-containing formulations and a control formulation containing PS80 (57).

All test formulations remained stable after multiple freeze/thaw cycles except for formulations (2), (4), (19), (22) and (26). These formulations contained high concentrations of mannitol as the only stabilizing excipient and showed a significant increase in aggregation after freezing.

After storage at 40°C for 1 week and after mechanical stress by orbital shaking and overhead rotation, the same trends as above were observed for formulations (1) to (57).

4. Detection of Acidic and Basic Species by IEX-HPLC Samples from the accelerated degradation study were diluted with eluent A (20 mM MES, pH 6.2) to a final concentration of 1 mg/mL and 30 μL was injected onto a MabPac SCX-10 (Thermo Scientific, 4×250 mm, 10 μm) column to detect the modification of pembrolizumab resulting in charge heterogeneity. Proteins were eluted by applying a salt gradient (KCl) with a mixture of mobile phase A (20 mM MES, pH 6.2) and mobile phase B (20 mM MES, 120 mM KCl, pH 6.2) at a flow rate of 1 mL/min at 40° C.

The eluted species were detected at a wavelength of 280 nm and displayed on a graph showing the concentration of the eluted species versus time. The elution profile showed a main peak as well as several peaks representing acidic and basic variants. The areas of all peaks were determined.

Table 4 summarizes the percentage of peak area of acidic species relative to the total peak area of eluting species for the formulations shown in Table 1. Each sample was measured in triplicate.
Table 4: Summary of basic species determined using IEX-HPLC

After 2 weeks of storage at 40° C., the levels of acidic species increased by 19% to 30% compared to t0 (17%) for the formulations tested in the design of experimental set-up.
For formulations (1)-(32), the effect of the buffer can be summarized as follows: histidine has a strong stabilizing effect, and for citrate, a slight destabilizing effect was observed at high temperatures, which decreased with decreasing pH. Therefore, histidine and citrate (especially at low pH) are the optimal buffer system for pembrolizumab.

For formulations (33)-(56), citrate buffer was used as the core component.
Comparing formulations (33)-(56) containing amino acids and/or glycerol as stabilizing excipients, optimal conditions for pembrolizumab formulation were identified in samples (36), (38), (39), (45), (47), (48) and (54). Further evaluation revealed that in terms of acidic species levels, the combination of high concentrations of arginine and/or lysine in the citric acid-based formulations with 100 mM glycerol could achieve results comparable to the sugars and/or sugar alcohols used in the histidine-based formulations (1)-(32). Formulation (56) containing poloxamer 188 provided results comparable to the polysorbate 20-containing formulation and the control formulation containing polysorbate 80 (57).

After storage at 40°C for 1 week, after multiple freeze/thaw cycles, and after mechanical stress via orbital shaking and overhead rotation, the same trends as above were observed for formulations (1) to (57).

In addition, for basic species, the same trends as for acidic species were observed.
5. Summary of Study Results Based on the results obtained using the presented method set, several excipients were identified that stabilized pembrolizumab in the liquid formulation and were superior to other excipients tested or compared to the reference product formulation. The formulation conditions and the influence of each factor that could have a stabilizing effect are shown in Figures 1 to 16.

We found that trehalose stabilized pembrolizumab over a very wide concentration range against aggregation at high temperatures (40°C) as well as after multiple freeze/thaw cycles, whereas formulations containing mannitol showed increased levels of aggregation after freeze/thaw stress.

In addition, the combination of high concentrations (100-300 mM) of arginine and/or lysine with 100 mM glycerol is expected to have a stabilizing effect against chemical modifications such as those that lead to aggregation and charge heterogeneity of pembrolizumab. Thus, a pembrolizumab formulation without any sugars or sugar alcohols appears feasible. The other two amino acids tested (glycine and proline) had equivocal and small effects on pembrolizumab stability. Therefore, further development of the test formulation will focus on arginine, lysine and/or glycerol.

Histidine and low molar concentrations (5-10 mM) of citrate significantly stabilize pembrolizumab. The preferred pH of the histidine and citrate buffer was identified as pH 5.5. Citrate-containing formulations were also stable at lower pH values down to pH 5.0.

The use of poloxamer 188 as a surfactant stabilized pembrolizumab formulations to the same extent as the use of polysorbate 20 and polysorbate 80 (control formulations), respectively.

Example 2: Formulations selected for stability testing Based on the results shown in Example 1, ten formulations were selected for testing in short-term stability testing. Using the same starting materials as described in Example 1, the formulations were prepared by dialysis. In addition, pembrolizumab in Keytruda®-labeled formulation buffer was included in the study as a control sample (11), which was directly dialyzed together with the other formulations. The ten different formulations (plus control sample) are shown in Table 5.

Table 5: Detailed information on the formulations prepared in this study (Example 2)

Samples are stored for up to 24 months under the conditions shown in Table 6. In addition, samples are subjected to two conditions of mechanical stress as well as five freeze/thaw cycles.
Table 6. Storage stability test of pembrolizumab

Protein stability is determined by size exclusion chromatography (SE-HPLC) for the presence of high molecular weight species (HMWS) and by SDS-cGE (non-reduced) for the presence of low molecular weight species (LMWS) and HMWS. Chemical modifications such as glycation, oxidation and deamidation are quantified by LC-ESI-MS and MS/MS in reduced peptide mapping. Ion exchange chromatography (IEX-HPLC) and imaging capillary isoelectric focusing (icIEF) are used to detect modifications that result in charge heterogeneity. In addition, samples are analyzed for appearance, turbidity, subvisible particle content and particle size. Protein concentration of samples is determined by UV-VIS spectroscopy. pH of formulations is measured only at t0.

Example 3: Formulations tested in stability studies 1. Sample preparation Based on the results obtained in Example 1, the six best performing formulations were selected and tested.

Using 25 mg/mL pembrolizumab as the starting material, seven formulations listed in Table 5 (six alternative formulations (1)-(6) and reference formulation (7)) were prepared by three-step dialysis. Pembrolizumab in Keytruda® labeled formulation buffer was included in the study as a control sample (7) and was directly dialyzed with the other formulations shown in Table 5. The dialyzed pembrolizumab was adjusted to 25 mg/mL ± 20% and sterile filtered at 0.22 μm using a PES membrane. The formulations shown in Table 5 were first analyzed for protein concentration by UV-VIS spectroscopy at 280 nm and precise pH adjustment was performed before initiating storage stability.

Table 7: Detailed information on the formulations prepared in this study

Storage conditions are shown in Table 8. Briefly, samples were stored at 5°C and 25°C/60% relative humidity (RH) for up to 3 months and at 40°C/75% RH for up to 1 month. Additionally, samples were subjected to two conditions of mechanical stress (overhead rotation, orbital shaking) and five freeze/thaw cycles (-80°C/+25°C). Sample storage continues for up to 24 months as listed in Table 8.

Table 8: Pembrolizumab storage conditions/stability program

Protein stability was determined by size exclusion chromatography (SE-HPLC) for the presence of high molecular weight species (HMWS) and by SDS-cGE (non-reduced) for the presence of low molecular weight species (LMWS) and HMWS. Chemical modifications such as glycation, oxidation and deamidation were quantified by LC-ESI-MS and MS/MS in reduced peptide mapping. Ion exchange chromatography (IEX-HPLC) and imaging capillary isoelectric focusing (icIEF) were used to detect modifications resulting in charge heterogeneity. In addition, samples were analyzed for appearance, turbidity, subvisible particle content and particle size. Protein concentration of samples was determined by UV-VIS spectroscopy. pH of formulations was measured at t0, 1 month, 3 months, 6 months, 9 months and 12 months.

2. Analysis of protein content by UV-VIS, measurement of pH, visible particles, sub-visible particles and osmolality.
All prepared formulations according to Table 7 were analyzed for their protein content as well as pH and osmolality before starting the stability study (t0). All samples of formulations according to Table 7 met the acceptance criteria for protein content (25 mg/mL ± 10% pembrolizumab) and pH (target ± 0.2). No visible particles or color changes were detected in all samples.

pH was analyzed using a SevenExcellence Multiparameter system and an InLab Micro Pro-ISM pH electrode (both from Mettler Toledo). Measurements were performed at 23°C-25°C according to USP using 150 μl of solution filled in 0.5 mL vials. For protein content measurements, a NanoPhotometer N120 from Implen was used, where 2 μl of sample solutions were diluted 1:10 with their relevant placebo buffers and quantified at 280 nm using an extinction coefficient of 1.418 l/g*cm. Background correction was performed using a wavelength of 320 nm.

During the stability program, all samples did not change in their solution pH, and all samples met the 5.5 ± 0.2 criterion at all storage conditions, even when using citrate buffer at the lower level of 5 mM. Thus, lowering the citrate concentration sufficiently stabilized the pH over 9 months. pH was determined in a single measurement.

Additionally, pembrolizumab concentrations were stable throughout the full stability program, with all results quantified within the acceptance criteria of 25 mg/mL ± 10% pembrolizumab. Pembrolizumab concentrations were determined in duplicate.

When the different formulations were analyzed for their visual appearance, they all showed identical results during the stability study when the formulations were compared with each other for one storage condition. At t0, all samples were analyzed to be clear and transparent. After physical stresses such as freeze/thaw and shaking/rotation, all formulations were analyzed to be clear with very slight turbidity. After 9 months of incubation at 5°C and 3 months of incubation at 25°C, respectively, all formulations were inspected to be clear with slight opalescence, meeting the specifications for pembrolizumab. For visual inspection, samples were analyzed in duplicate.

A FlowCam 8100 Multi Objective from Anasysta was used to analyze particles subvisible to the naked eye. These micron-sized protein aggregates and particles are an important quality attribute of therapeutic protein formulations due to the risk of enhancing immunogenic responses. The method was tuned to the following parameters: analysis efficiency: 60-70%; auto image range: 30 fps; distance to nearest neighbor: 3 μm; flow rate: 0.150 mL/min; sample volume processed: 0.100 mL.

All formulations performed equally well after mechanical stress, freeze/thaw stress, and after incubation at 5°C for 9 or 12 months, 25°C for 3 months, and 40°C for 1 month, falling within the USP <788> pharmacopoeia specifications. Formulations (1) and (2) using arginine as a stabilizer had the lowest number of particles in the 10 μm or larger size range after 12 months storage at 5°C, followed by a slight increase in the number of lysine-containing formulation (4) and sugar-containing formulations (2) and (7). Only a very slight increase in all particles with a size below 25 μm was observed after 1 month storage at 40°C. Results for the other storage conditions were consistent within the method variability. All measurements were performed in duplicate to analyze particles subvisible to the naked eye by FlowCam.

3. Analysis of high molecular weight species (HMWS) by SE-HPLC.
Test samples were diluted with 20 mM histidine, pH 5.5, to a final concentration of 1 mg/mL, and 5 μL was injected onto a TSKgel UP-SW3000 (Tosoh, 4.6×150 mm, 2 μm) column to detect high molecular weight species (HMWS) of pembrolizumab.

Proteins were eluted by isocratic elution using 0.1 M sodium phosphate buffer (pH 6.7) containing 0.05% (v/v) sodium azide at a flow rate of 0.35 mL/min at 25°C. Eluting species were detected at a wavelength of 280 nm and displayed on a graph showing the concentration of eluting species versus time. The elution profile showed a monomer peak with non-aggregated protein and multiple peaks of protein representing high molecular weight species (HMWS) of protein. The areas of all peaks were determined.

Table 9 summarizes the percentage of HMWS peak area relative to the total peak area of eluting species for the formulations shown in Table 7. Sample acquisition began after 6 months to narrow the formulation set to the most promising candidates, at which point incubation of all remaining formulations was stopped. Each sample was measured in triplicate.

Table 9: Summary of HMWS determined using SE-HPLC

After storage for up to 9 months at 5°C (target storage conditions), HMWS remained at t0 levels for all formulations tested in this stability study, and long-term storage stability at these conditions is expected. Slightly better results were achieved by using formulations (1) and (2) containing arginine as a stabilizer. After storage for up to 12 months at 5°C (target storage conditions), slightly better results were achieved by using formulations (1) and (2) containing arginine as a stabilizer, and formulation (4) containing lysine. After storage for up to 3 months at 25°C, HMWS levels increased slightly to 0.9% to 1.2% compared to t0 (0.8 and 0.9%, respectively). The lowest HMWS levels after storage for up to 3 months at 25°C were observed for formulations (1), (2) and (6) containing the amino acid arginine (arginine alone or in combination with lysine), followed by formulation (4) containing only lysine. Trehalose-containing formulations (3) and (5) showed a slight increase in HMWS levels after up to 3 months of storage at 25° C., but surprisingly no relevant increase in HMWS was detected at the target storage conditions. In addition, all formulations tested remained stable after multiple freeze/thaw (F/T) cycles and after mechanical stress. Overall, all formulations tested in this study showed similar or improved results compared to the reference formulation (7).

4. Detection of Acidic and Basic Species by IEX-HPLC The test samples were diluted with eluent A (20 mM MES, pH 6.2) to a final concentration of 1 mg/mL and 30 μL was injected onto a MabPac SCX-10 (Thermo Scientific, 4×250 mm, 10 μm) column to detect the modification of pembrolizumab that results in charge heterogeneity. Proteins were eluted by applying a salt gradient (KCl) with a mixture of mobile phase A (20 mM MES, pH 6.2) and mobile phase B (20 mM MES, 120 mM KCl, pH 6.2) at a flow rate of 1 mL/min at 40° C.

The eluted species were detected at a wavelength of 280 nm and displayed on a graph showing the concentration of the eluted species versus time. The elution profile showed a main peak as well as several peaks representing acidic and basic variants. The areas of all peaks were determined.

Table 10 summarizes the percentage of peak area of acidic species relative to the total peak area of eluting species for the formulations shown in Table 7. Each sample was measured in triplicate.
Table 10: Summary of basic species determined using IEX-HPLC

After storage at 5°C (target storage conditions) for up to 3 months, both acidic and basic species remained at t0 levels for all formulations tested in this stability study, indicating long-term stability. After storage at 5°C (target storage conditions) for up to 9 months, both acidic and basic species only increased slightly. After storage at 5°C (target storage conditions) for up to 12 months, both acidic and basic species remained at t0 levels for all formulations tested in this stability study, indicating long-term stability, with the lowest variability compared to t0 shown in formulations (1), (2) and (4) containing one or two amino acids (either arginine or lysine, or both arginine and lysine). After storage at 25°C for up to 3 months, acidic species levels increased slightly to 20.1% to 23.9% compared to t0 (approximately 17.5%). The lowest acidic species levels after storage were observed for formulations (1), (2), (4) and (6) containing one or two amino acids (either arginine or lysine, or both arginine and lysine). Trehalose-containing formulations (3) and (5) showed only a slight increase in acidic species levels, which is comparable to the reference formulation (7). In addition, all formulations tested remained stable after multiple freeze/thaw cycles and after mechanical stress.

The same trend was observed for the generation of basic species. After storage at 25°C for up to 3 months, the basic species level was slightly reduced to 23.8%-29.0% compared to t0 (30.4%-30.6%). The best results (here the highest content) of basic species level after storage were again observed for formulations (1), (2), (4) and (6) containing one or two amino acids (either arginine or lysine, or both arginine and lysine). Trehalose-containing formulations (3) and (5) showed only a slight reduction in basic species level, which is comparable to the reference formulation (7). Overall, all formulations tested in this study showed similar or better results compared to the reference formulation (7).

5. Analysis of low molecular weight species (LMWS) by SDS-cGE (non-reducing).
Capillary gel electrophoresis for quantification of LMWS was performed using the LabChip GX II Touch Protein Characterization System and Protein Express Assay Kit (catalog no. CLS960008) (both Perkin Elmer).

To prepare the sample denaturing solution, 5400 μl of Protein Express sample buffer from the assay kit was mixed with 300 μl of 10% LDS (lithium dodecyl sulfate dissolved in water) and 300 μl of 200 mmol/L NEM solution (N-ethylmaleimide dissolved in water). 7 μl of this denaturing solution was mixed with 2 μl of pembrolizumab-containing sample (prediluted to 2 mg/mL in water). For denaturation, the samples were incubated at 75°C for 10 min. Then, 35 μl of water was added to each sample immediately before analysis. Analysis was performed by using the assay preset from the method "P200 Antibody Analysis" of the Perkin Elmer instrument software.

For cGE, separations were performed by forward injection into a neutral bare fused silica capillary (20 cm effective length, 50 μm diameter) using a Beckman Coulter PA800 plus instrument. Protein separation was performed by applying a voltage (5 kV for 20 s) to the capillary followed by 15 kV for 30 min for reducing conditions and 15 kV for 40 min for non-reducing conditions. UV absorption was measured at 220 nm using a PDA detector and a 100 x 200 aperture. The capillary temperature was kept constant at 25°C for all steps. The autosampler temperature was set at 15°C. Data were evaluated for peak integration using 32Karat software (Beckman Coulter). Peak areas were determined as velocity-corrected relative peak areas, taking into account the fact that in capillary electrophoresis, early peaks migrate through the detector window faster than late peaks. Sample peak integration was performed relative to the electropherograms of formulation buffer or pure water blanks to identify and exclude non-protein specific peaks.

Table 11 summarizes the percentage of LMWS peak area relative to the total peak area of the eluting species for the formulations shown in Table 7. Each sample was measured in triplicate.
Table 11: Summary of LMWS determined using SDS-cGE (non-reduced)

After storage at 5°C (target storage conditions) for up to 3 months, LMWS increased slightly to about 0.7% compared to t0 (about 0.4%) for all formulations tested in this stability study, predicting successful long-term storage stability. Essentially the same observations were made after storage at 5°C for 9 months. After storage at 5°C (target storage conditions) for up to 12 months, LMWS increased slightly to about 0.6% to about 0.7% compared to t0 (about 0.4%) for all formulations tested in this stability study, predicting successful long-term storage stability. After storage at 25°C for up to 3 months, LMWS levels increased to about 1.1% compared to t0 (about 0.4%), with no significant differences between the tested formulations. In addition, all formulations remained stable after multiple freeze/thaw cycles and after mechanical stress. Overall, all formulations tested in this study showed similar very good results, as well as compared to the reference formulation (7).

6. Detection of Modifications by LC-ESI-MS and -MS/MS Analysis by LC-ESI-MS and -MS/MS was used for sequencing of pembrolizumab by peptide mapping. Several post-translational modifications (oxidation, deamidation and glycation) were quantified using several different combinations of digestion conditions followed by LC-ESI-MS and -MS/MS measurements. LC-ESI-MS and -MS/MS mass spectra were obtained using an UlTiMate® 3000 system (Thermo Fisher Scientific) coupled to a Q Exactive Orbitrap Plus mass spectrometer (Thermo Fisher Scientific). Separation of peptides was performed by reversed-phase (RP) chromatography on an Accucore RP-MS LC column (2.1 x 100 mm, 2.6 μm particle size, Thermo Fisher Scientific). The following eluents were used: A: water with 0.1% formic acid; B: acetonitrile with 0.1% formic acid. A segmented gradient was applied from 3% B to 36% B in 45 min at 30°C with a flow rate of 0.4 mL/min. MS and MS/MS spectra (generated using higher energy collisional dissociation (HCD)) were recorded in positive ion mode with internal mass calibration. The dataset was searched using Protein Metrics Byonic™ against a sequence database representing the pembrolizumab sequence and common contaminants (e.g., sequences of proteases used in digestion) and analyzed for asparagine/glutamine deamidation levels, methionine/tryptophan oxidation levels, and glycation levels. Samples from pull points of the same stability were simultaneously proteolytically digested, reduced, and alkylated.

Samples shown in Table 7 were analyzed in a single measurement before and after one month of incubation at 40°C/75% relative humidity. The pull point after one month of incubation at 40°C was chosen to evaluate trends at accelerated conditions. Accelerated conditions also occur at the target storage temperature, but at a slower rate. After three months at 5°C, modifications will likely not exceed method variability. Therefore, analysis of the later pull point is more relevant for this storage condition.

Table 12 summarizes the deamidation levels, Table 13 summarizes the oxidation levels, and Table 14 summarizes the glycation levels for the formulations shown in Table 1.
Table 12: Summary of deamidation levels determined using LC-ESI-MS and -MS/MS

Table 13: Summary of oxidation levels determined using LC-ESI-MS and -MS/MS

Table 14: Summary of glycation levels determined using LC-ESI-MS and -MS/MS

Total deamidation levels increased significantly after storage at 40°C for 1 month. The lowest deamidation content was obtained using formulations (1) and (2), both of which are based on the use of arginine. All other formulations resulted in higher levels of total deamidation after storage.

Total oxidation levels increased after storage at 40°C for 1 month. Slightly smaller increases were observed in trehalose-containing formulations (3) and (5) and in formulations using arginine without lysine, i.e., formulations (1) and (2). The highest oxidation levels were observed in lysine-containing formulations (4) and (6) and the reference formulation (7).

Glycation levels remained within method variability after 1 month of storage at 40°C and were not significantly different between the alternative formulations. In contrast, the reference formulation (7), which uses sucrose as an excipient, showed higher glycation levels after storage compared to all alternative formulations.

7. Summary of Study Results Based on the results obtained in this stability program analyzed using the method sets shown, novel compositions were identified that were able to stabilize pembrolizumab in liquid formulations and were superior compared to other excipients tested or compared to the reference product formulation.

In particular, the use of arginine improved the stability of 25 mg/mL pembrolizumab. Formulations (1) and (2) containing 300 mM arginine, 0.2 mg/ml polysorbate 80, 5 mmol/L or 10 mmol/L citric acid at pH 5.5 provided very promising results, especially with regard to the chemical stability of pembrolizumab. For example, IEX-HPLC showed the best stability of pembrolizumab in formulations (1) and (2) with respect to acidic and basic species, and deamidation analyzed by peptide mapping was also the lowest in formulations (1) and (2) compared to all other tested formulations. A similar effect was shown when arginine was replaced by the amino acid lysine (4) or mixed with lysine (6). Also, the formation of HMWS was lowest with formulations (1) and (2), both of which are based on the use of arginine as a stabilizer.

However, the use of trehalose in formulations (3) and (5) also yielded encouraging stability data when the drug product was stored at the target storage temperature of 2°C to 8°C, where all relevant stability data were stable and comparable to the other test formulations, including the reference formulation. In any case, the use of the reference formulation resulted in more undesired glycation when compared to the other formulations due to the presence of sucrose.

The use of a citric acid/sodium citrate buffer system showed a slight advantage when the concentration was reduced to 5 mM, especially in combination with trehalose. Also, reducing the buffer concentration or buffering capacity at pH 5.5, respectively, successfully stabilized the pH during the stability program.

II. Formulation Development of Highly Concentrated Pembrolizumab Formulations Example 1: Sample Preparation of Highly Concentrated Pembrolizumab in Histidine Buffer A desired therapeutic monoclonal antibody (mAb) formulation was prepared using an ultrafiltration/diafiltration (UF/DF) procedure. Here, pembrolizumab was transferred by UF/DF and subsequent addition of polysorbate 20 into a formulation containing 10 mM L-histidine/histidine HCl, 205 mM trehalose dihydrate, 0.01% (w/v) polysorbate 20 (pH 5.5) to 100 mg/mL or 150 mg/ml of pembrolizumab by the following steps: 13 grams of pembrolizumab at a concentration of 11.4 mg/mL was concentrated by UF/DF and transferred into a formulation containing 10 mM L-histidine/histidine HCl, 205 mM trehalose dihydrate (pH 5.5) to a target concentration of 120 mg/mL.

In this step, a Pellicon® 3 Cassette with Biomax® 30 kDa Membrane, D screen was used (Merck Millipore) to provide a membrane load of approximately 13 g pembrolizumab/264 ^cm2 . The process conditions were a transmembrane pressure of 1 bar with a feed flow rate of 6 L/ ^m2 /min. Buffer exchange was performed within 5 diafiltration volumes using formulation buffer without polysorbate 20.

The solution, now containing 120 mg/mL pembrolizumab, was then sterile filtered without pre-cleaning the filter by using a Sartopore 2 XLG 0.22 μm PES filter (filter area 210 cm ² ). The final concentration of pembrolizumab was quantified to be 116.9 mg/mL, which was a good process recovery of 91.6%.

The solution was then split 50:50. The first portion was diluted with 10 mM L-histidine/histidine HCl, 205 mM trehalose dihydrate (pH 5.5) to a final concentration of 100 mg/mL pembrolizumab in 10 mM L-histidine/histidine HCl, 205 mM trehalose dihydrate (pH 5.5) and polysorbate 20 was added to a final concentration of 0.01% (w/v) polysorbate 20 in the solution.

The second portion was transferred to a Vivaspin filter (Sartorius Stedim) using a 30 kDa PES membrane, successfully increasing the concentration of pembrolizumab to 170 mg/mL. The concentration of pembrolizumab was then adjusted to 150 mg/mL by dilution with 10 mM L-histidine/histidine HCl, 205 mM trehalose dihydrate (pH 5.5), and then sterile filtered using a bottle-top vacuum filter (PES filter membrane area: 13.6 μm) with a process recovery of 95%. Finally, polysorbate 20 was added to the formulation to a final concentration of 0.01% (w/v).

Table 15 shows the analytical results covering the different steps during the preparation of samples containing pembrolizumab concentrations of approximately 100 mg/mL or 150 mg/mL and the final processed samples. The processing was very gentle and smooth, with HMWS determined by SE-HPLC showing only a slight increase from 0.84% HMWS to 1.10% in the 100 mg/mL pembrolizumab sample and a comparable 1.08% in the 150 mg/mL pembrolizumab sample. Other impurities such as LMWS (quantified by CE-SDS (non-reduced)), acidic and basic species (quantified by HP-CEX) were also within acceptable ranges and were not altered by the processing steps. With a well sterile filtered solution through a 0.22 μm filter, there was negligible loss of material, making pembrolizumab in the described histidine buffer a very good formulation candidate suitable for achieving high protein concentrations in a scalable manufacturing process. All methods are described in Example II.4.

Table 15: Preparation of high-concentration pembrolizumab by UF/DF and sample characterization

Example 2: Freeze/thaw stability of pembrolizumab at concentrations of 100 mg/mL or 150 mg/mL in histidine buffer Samples produced in Example 1 and shown in Table 16 were tested for stability to freeze/thaw stress and analyzed for the presence of HMWS by SE-HPLC (method described in Example 4).

Table 16: Detailed information on the formulations used in this study

For this purpose, 500 μl aliquots of both samples (a) and (b) were analyzed by SE-HPLC before and after the freezing process to below -65°C (according to the method described in Example 3).

Table 17: Summary of HMWS determined by SE-HPLC before and after the freeze/thaw process

Both concentrations of pembrolizumab 100 mg/mL (a) and 150 mg/mL (b) in 10 mM L-histidine/histidine HCl, 205 mM trehalose dihydrate, 0.01% polysorbate 20 (pH of solution 5.5) are stable to freeze/thaw stress. Both products can be frozen without increasing aggregation levels, which is a prerequisite for long-term storage of e.g. drug substance.

Example 3: Osmolality of 100 mg/mL or 150 mg/mL Pembrolizumab Concentration in Histidine Buffer Samples (a) and (b) prepared in Example 1 and shown in Table 16 were tested for osmolality. The goal of this development is a pembrolizumab formulation suitable for both intravenous and subcutaneous administration. Formulations for the intravenous route of administration of pembrolizumab can contain a wider range of osmolality, but solutions injected subcutaneously should be isotonic in the best case. Osmolality was determined in duplicate by freezing point depression using an Osmomat 3000 from Gonotec (Germany).

Table 18: Osmolality of pembrolizumab 100 mg/mL (a) and 150 mg/mL (b) in 10 mM L-histidine/histidine HCl, 205 mM trehalose dihydrate, 0.01% polysorbate 20 (solution pH 5.5)

Osmolality data indicates that both the 100 mg/ml and 150 mg/ml samples are suitable for subcutaneous administration.
Example 4: Real-time and accelerated stability studies of pembrolizumab concentrations of 100 mg/ml and 150 mL in histidine buffer Stability studies were performed with samples (a) and (b) according to Table 16. Samples were stored at -70°C, 5°C (target storage temperature), 25°C/60% relative humidity, and 40°C/75% relative humidity for up to 3 months.

Table 19: Detailed information on the stability program

Protein stability was determined by size exclusion chromatography (SE-HPLC) for the presence of high molecular weight species (HMWS) and by SDS-cGE (non-reducing) for the presence of fragments (LMWS) and HMWS. Imaging capillary isoelectric focusing (icIEF) was used to detect modifications resulting in charge heterogeneity.

Analysis of high molecular weight species (HMWS) by SE-HPLC.
Protein samples for stability studies were diluted to a concentration of 2.5 mg/mL with the corresponding formulation buffer and 20 μl of the dilution was applied to a TSKgl G3000SWXL (Tosoh, 300×7.8 mm, 5 μm) column to detect high molecular weight species of pembrolizumab.

Proteins were eluted by isocratic elution using 0.1 M potassium phosphate buffer (pH 5.6) containing 0.25 M potassium chloride at a flow rate of 0.5 mL/min at 25°C. Eluting species were detected at a wavelength of 280 nm and displayed on a graph showing the concentration of eluting species versus time. The elution profile showed a major peak with non-aggregated protein and multiple peaks of protein representing higher molecular weight forms of the protein. The areas of all peaks were determined.

Table 20 shows the percentage of HMWS peak area relative to the total peak area of eluting species for the stability study samples shown in Table 19. Each sample was tested in duplicate.

Table 20: Summary of HMWS determined by SE-HPLC

At the target storage condition of 5°C, the formation of HMWS is very low and the product is stable against aggregate formation. This is valid at both tested concentrations for the formulations containing 100 mg/mL pembrolizumab and 150 mg/mL pembrolizumab. After 3 months of storage, HMWS increased slightly compared to the starting point, from 0.73% to 0.96% for the 100 mg/mL sample and from 0.89% to 1.13% for the 150 mg/mL sample. In addition to real-time storage, accelerated conditions also provided very encouraging stability results, as HMWS was quantified in an acceptable range for stressed storage.

When samples were stored frozen at -70°C, the 100 mg/mL sample showed no increase in HMWS after 3 months, and the 150 mg/mL sample showed only a slight increase to 1.55% HMWS.

Detection of HMWS and LMWS by SDS-cGE (non-reduced) Capillary gel electrophoresis was performed based on the IgG Purity and Heterogeneity Analysis established by Beckman Coulter. For non-reduced analysis, samples were diluted to a final concentration of 1 mg/mL in SDS sample buffer pH 6.4-6.9 (10 mM phosphate citrate, 1% SDS). The thiol alkylating reagent N-ethylmaleimide (NEM; 10 mM) was then added to the sample mixture to prevent fragmentation. Prior to analysis, samples were heat denatured at 70°C for 10 min. For the cGE method, separation was performed by forward injection into a neutral bare fused silica capillary (20 cm effective length, 50 μm diameter) using a Beckman Coulter PA800 plus instrument. Protein separation was performed by applying a voltage (5 kV for 20 seconds) to the capillary followed by 15 kV for 30 minutes for reducing conditions and 15 kV for 40 minutes for non-reducing conditions. UV absorption was measured at 220 nm using a PDA detector and 100 x 200 aperture. The capillary temperature was kept constant at 25°C for all steps. The autosampler temperature was set at 15°C. Data was evaluated for peak integration using 32Karat software (Beckman Coulter). Peak areas were determined as velocity-corrected relative peak areas, taking into account the fact that in capillary electrophoresis, early peaks migrate through the detector window faster than late peaks. Sample peak integration was performed by comparison with electropherograms of formulation buffer or pure water blanks to identify and exclude non-protein specific peaks.

The samples shown in Table 16 were incubated at 5° C. or 40° C./75% relative humidity for 1 month or 3 months, frozen at −70° C., and then analyzed in a single measurement.
According to Table 21, both test formulations (containing pembrolizumab at 100 mg/mL and 150 mg/mL) were stable upon storage for 3 months at the target storage temperature of 5° C., with LMWS showing only a slight increase over this period, predicting long-term stability in terms of LMWS formation as tested by SDS-cGE (non-reduced) (1.105% to 1.193% at 100 mg/mL; 1.116% to 1.170% at 150 mg/mL). Negligible HMWS was detected in both formulations during this incubation period (0.090% to 0.151% at 100 mg/mL; 0.139% to 0.166% at 150 mg/mL). Additionally, no or very little LMWS or HMWS was formed when stored frozen at −70° C.

Storage at higher temperatures (40°C/75% RH for 1 month) resulted in slightly increased values as expected for storage at accelerated conditions, with both test formulations containing LMWS and slightly increased HMWS in very similar ranges.

Table 21: Summary of LMWS and HMWS determined by SDS-cGE (non-reducing)

Detection of Acidic and Basic Species by icIEF The goal of imaging capillary isoelectric focusing (icIEF) is to determine the heterogeneity of protein isoelectric points (pI) and charge isoforms caused by post-translational modifications (PTMs). The power of icIEF lies in the high-resolution electropherogram that allows accurate and reproducible relative quantification of charge isoforms.

Samples were rebuffered by ultrafiltration against ultrapure water and interfering buffer components were removed from the samples using Detergent Removal Spin Columns (Pierce) according to the manufacturer's instructions. Protein concentration was determined by UV measurement (absorbance at 280 nm). Rebuffered samples were diluted to 1 g/L with ultrapure water. As indicated, 40 μg of diluted sample (corresponding to a final protein concentration of 0.2 g/L) was mixed with 2% ampholytes (0.25% pH 9-11 and 3% 8-10.5), 0.35% methylcellulose, 4 M urea and pI markers (8.18 and 9.99). Electrophoresis was performed in two steps (1500 V for 1 min and 3000 V for 10 min). Final analysis was performed using the imaged CEsystem Maurice C (ProteinSimple). Data were evaluated by pI calibration of the electropherograms using two internal pI markers and Compass for iCE software (ProteinSimple). Peak integration was performed using Empower software (Waters). Calculations using relative peak areas (e.g., for peak grouping) were performed using initial values of relative peak areas to three decimal places. Initial values used in the calculations and the calculation results are reported rounded to one decimal place.

Samples were analyzed in one measurement during incubation at 5° C. and 40° C./75% relative humidity for 3 months.
According to Table 22, the 3-month data at 5° C. demonstrates excellent stability of pembrolizumab in both formulations (100 mg/mL and 150 mg/mL) as no significant changes in acidic and basic species were quantified. Also, no additional acidic or basic species were generated when stored frozen at −70° C. During storage at 40° C./75% relative humidity, a shift towards acidic species was detected, as expected for storage under accelerated conditions. This shift resulted in a decrease in the main peak and basic species in both formulations.

Table 22: Summary of acidic and basic species determined by cIEF

Example 5: Rheometric Viscosity Study of High Concentration Samples in Histidine Buffer The purpose of this example was to measure the dynamic viscosity of pembrolizumab in formulations with different antibody concentrations. As shown in Table 8, both concentrations of pembrolizumab in 10 mM L-histidine/histidine HCl, 205 mM trehalose dihydrate, 0.01% polysorbate 20 (pH of solution 5.5) were analyzed, as well as a 25 mg/mL sample in the same formulation prepared by diluting the 100 mg/mL sample 1:4 with the formulation.

The kinematic viscosity of the three samples was determined using a Kinexus ultra plus flowmeter at 20°C and a shear rate of 250 s-1. The method and cone/plate geometry according to Table 23 were applied to analyze the viscosity.

Table 23: Method settings for viscosity measurements

All samples were analyzed in triplicate and the results are shown in Table 16. Formulations containing 25 mg/mL and 100 mg/mL pembrolizumab showed very low dynamic viscosities of 1.463 mPa*s and 4.646 mPa*s, respectively. The 150 mg/mL pembrolizumab formulation showed an increased but still acceptable viscosity of 17.88 mPa*s, allowing for routine DSP processing, drug substance and drug product filling, and injectability by using standard prefilled syringes for subcutaneous administration.

Table 24: Dynamic viscosity obtained for three different concentrations of pembrolizumab

Example 6: Determination of break loose force and gliding force using prefilled syringes suitable for subcutaneous injection To test the suitability of the high concentration formulations for subcutaneous administration, 1 mL of both formulations according to Table 16 (containing 100 mg/mL and 150 mg/mL pembrolizumab) were filled into a glass syringe with a nominal volume of 1 mL with a 29 gauge staked needle (BD Neopak 1MILL 29G TW ^1/2 _5B RNS BD260, catalog no. 47432910). The syringe has a stopper attached to the vent tube (BD Hypak BSCF 1MLL W4023 Flur DAIKYO LID, catalog number 47284410).

After filling, the syringes were tested for injection force simulating injection at an injection rate of 190 mm/min by using a tensile tester (ZwickiLine 500N, ZwickRoell GmbH & Co. KG, Ulm, Germany). The use of the tensile machine allows recording of the applied force in a force-displacement plot. The injection force of the syringe is divided into the sliding yield stress and the sliding equilibrium stress. The sliding yield stress is the force required to initiate the movement of the plunger, and the sliding equilibrium stress (dynamic sliding equilibrium stress) represents the force required to maintain the continuous movement of the plunger.

The plunger rod was attached to the stopper without moving the stopper, and the system was allowed to equilibrate to room temperature before measurements were taken.
Table 25. Injection force of high concentration pembrolizumab in prefilled syringes

The results (see Table 25) show sliding yield stress and sliding equilibrium stress (average and maximum) of less than 15 N for both formulations containing 100 mg/mL and 150 mg/mL pembrolizumab when releasing the contents. These values allow for easy and safe subcutaneous injection of highly concentrated solutions into patients.

Example 7: Pembrolizumab at concentrations of 100 mg/mL and 150 mg/mL in a citrate-based alternative formulation Pembrolizumab is transferred by UF/DF to the citrate buffer formulation shown in Table 26 and concentrated to target concentrations of 100 mg/mL and 150 mg/mL of pembrolizumab.

Table 26: Detailed information on the products used in Example 7

After preparation, the samples are processed by freeze/thaw cycles. In addition, real-time (2°C-8°C) and accelerated stability tests (25°C, 40°C) are performed to analyze the stability of the products against the formation of HMWS by SE-HPLC. Viscosity tests of the samples are performed by using a flow meter and functionality by simulating subcutaneous injection using a pre-filled syringe is also tested by a tensile machine.

Example 8: Sample preparation of high concentration pembrolizumab in citrate buffer Pembrolizumab was transferred by UF/DF into the citrate buffer formulation shown in Table 27 and concentrated to target concentrations of 100 mg/mL and 150 mg/mL of pembrolizumab.

Table 27: Detailed information on the products used in Example 8

Pembrolizumab was transferred to 100 mg/mL or 150 mg/mL pembrolizumab by UF/DF and subsequent addition of polysorbate 80 into a formulation containing 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/mL polysorbate 80 (pH 5.5) by the following steps: 15 grams of pembrolizumab at a concentration of 10.8 mg/mL was concentrated by UF/DF and transferred into a formulation containing 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate (pH 5.5) to a target concentration of 120 mg/mL.

In this step, a Pellicon® 3 Cassette with Biomax® 30 kDa Membrane, D screen was used (Merck Millipore) to provide a membrane load of approximately 15 g pembrolizumab/264 ^cm2 . The process conditions were a transmembrane pressure of 1 bar with a feed flow rate of 6 L/ ^m2 /min. Buffer exchange was performed within 5x diafiltration volume with formulation buffer without polysorbate 80.

The resulting solution containing 121.6 mg/mL pembrolizumab was then sterile filtered without prior filter washing by using a Sartopore 2 XLG 0.22 μm PES filter (filter area 210 cm ² ). The final concentration of pembrolizumab was determined to be 119.8 mg/mL with a very good process recovery of 93.5% compared to the starting material containing 10.8 mg/mL pembrolizumab.

The solution was then split 50:50. The first portion was diluted with 10 mM L-citric acid/sodium citrate, 205 mM trehalose dihydrate (pH 5.5) to a final concentration of 100 mg/mL pembrolizumab in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate (solution pH 5.5), and polysorbate 80 was added to a final concentration of 0.2 mg/mL polysorbate 80 in the solution.

The second portion was transferred to a Vivaspin filter (Sartorius Stedim) using a 30 kDa PES membrane, successfully increasing the concentration of pembrolizumab to 161 mg/mL. The concentration of pembrolizumab was then adjusted to 150 mg/mL by dilution with 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate (pH 5.5), and then sterile filtered using a bottle-top vacuum filter (PES filter membrane area: 13.6 μm). Finally, polysorbate 80 was added to the formulation to a final concentration of 0.2 mg/mL.

Table 28 shows analytical results covering the different steps during the preparation of samples containing pembrolizumab concentrations of approximately 100 mg/mL or 150 mg/mL and the final processed samples. The processing was very gentle and smooth, with HMWS determined by SE-HPLC showing only a slight increase from 0.85% HMWS to 1.20% in the 100 mg/mL pembrolizumab sample and an increase to 1.31% in the 150 mg/mL pembrolizumab sample. Other impurities such as LMWS (quantified by CE-SDS (non-reduced)), acidic and basic species (quantified by HP-CEX) were also within acceptable ranges and were not altered by the processing steps. With a well sterile filtered solution through a 0.22 μm filter, there was negligible loss of material, making pembrolizumab in the described citrate buffer a very good formulation candidate suitable for achieving high protein concentrations in a scalable manufacturing process. All methods are described in Examples 4, 5, and 6.

Table 28: Preparation of high concentration pembrolizumab in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80 (solution pH 5.5) by UF/DF and sample characterization

Example 9: Osmolality of Pembrolizumab at a concentration of 100 mg/mL or 150 mg/mL in Citrate Buffer The osmolality of samples (c) and (d) prepared in Example 8 and shown in Table 27 was determined. The aim of this development is a pembrolizumab formulation suitable for both intravenous and subcutaneous administration. Formulations for the intravenous route of administration of pembrolizumab can contain a wider range of osmolality, but solutions injected subcutaneously should be isotonic in the best case. Osmolality was determined in duplicate by freezing point depression using an Osmomat 3000 from Gonotec (Germany).

Table 29: Osmolality of pembrolizumab 100 mg/mL (c) and 150 mg/mL (d) in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80 (pH of solution 5.5)

The osmolality data shown in Table 29 indicate that both the 100 mg/ml and 150 mg/ml samples, all formulated in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80 (solution pH 5.5), are highly suitable for subcutaneous administration.

Example 10: Real-time and accelerated stability studies of Pembrolizumab at concentrations of 100 mg/ml and 150 mL in citrate buffer Stability studies were performed with samples (c) and (d) all formulated in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80 (pH of solution 5.5) according to Table 30. Samples were stored at -70°C, 5°C (target storage temperature), 25°C/60% relative humidity and 40°C/75% relative humidity for up to 3 months.

Table 30: Detailed information on the stability program

Protein stability was determined by size exclusion chromatography (SE-HPLC) for the presence of high molecular weight species (HMWS) and by SDS-cGE (non-reducing) for the presence of fragments (LMWS) and HMWS. Imaging capillary isoelectric focusing (icIEF) was used to detect modifications resulting in charge heterogeneity.

All procedures were carried out according to Example 4.
Analysis of high molecular weight species (HMWS) by SE-HPLC.
Table 31 shows the peak area percentage of the HMWS and main peak relative to the total peak area of the eluting species for the stability study samples shown in Table 30. Each sample was tested in duplicate.

Table 31: Summary of HMWS and LMWS determined by SE-HPLC

At the target storage condition of 5°C, the generation of HMWS is very low and the product is stable against aggregate formation. This is valid at both tested concentrations for the formulations containing 100 mg/mL pembrolizumab and 150 mg/mL pembrolizumab. After 3 months of storage, HMWS increased slightly compared to the starting point, from 0.79% to 1.07% for the 100 mg/mL sample and from 0.80% to 1.33% for the 150 mg/mL sample. In addition to real-time storage, accelerated conditions also provided very encouraging stability results, as HMWS was quantified in an acceptable range for stressed storage.

When samples were stored frozen at -70°C, the 100 mg/mL and 150 mg/mL samples showed no increase in HMWS after 3 months, a result that is within the variability of the assay. Both products can be frozen without increasing aggregation levels, which is a prerequisite for long-term storage of, for example, drug substances.

Detection of HMWS and LMWS by SDS-cGE (non-reducing) Samples (c) and (d) shown in Table 27 were analyzed in a single run after incubation at 5° C. or 40° C./75% relative humidity for 1 month or 3 months, or after freezing at −70° C. for 3 months.

According to Table 32, both test formulations (containing pembrolizumab at 100 mg/mL and 150 mg/mL) were stable at the target storage temperature of 5°C for 3 months, and LMWS showed no or only a slight increase during this period, predicting long-term stability with respect to LMWS formation as tested by SDS-cGE (non-reduced) (100 mg/mL: no increase; 150 mg/mL: 1.004% to 1.144%). Slightly increasing amounts of HMWS were detected in both formulations during this incubation period (100 mg/mL: 0.068% to 0.312%; 150 mg/mL: 0.106% to 0.450%). Also, no or only little LMWS or HMWS was formed when stored frozen at -70°C.

Storage at higher temperatures (1 and 3 months at 40°C/75% RH) increased values as expected for storage at accelerated conditions. Both test formulations contained LMWS and slightly increased HMWS in very similar ranges.

Table 32: Summary of LMWS and HMWS determined by SDS-cGE (non-reducing)

Detection of acidic and basic species by icIEF Samples according to Table 27 were analyzed in a single measurement after incubation at 5° C. or 40° C./75% relative humidity for 1 month or 3 months, or after freezing at −70° C. for 3 months.

According to Table 33, the 3-month data at 5°C demonstrates excellent stability of pembrolizumab in both formulations (100 mg/mL and 150 mg/mL) as no significant changes in acidic and basic species were quantified. Also, no additional acidic or basic species were generated when stored frozen at -70°C. During storage at 40°C/75% relative humidity, a shift towards acidic species was detected, as expected for storage under accelerated conditions. This shift resulted in a decrease in the main peak and basic species in both formulations.

Table 33: Summary of acidic and basic species determined by cIEF

Example 5: Rheometric Viscosity Study of High Concentration Samples in Citrate Buffer The purpose of this example was to measure the dynamic viscosity of pembrolizumab in formulations with different antibody concentrations. Both concentrations of pembrolizumab were analyzed in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80 (pH of solution 5.5), as shown in Table 27.

The kinematic viscosity of the two samples was determined using a Kinexus ultra plus flowmeter at 20°C and a shear rate of 250 s-1. The method and cone/plate geometry according to Table 34 were applied to analyze the viscosity.

Table 34: Method settings for viscosity measurements

All samples from Table 27 formulated in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80 (pH of solution 5.5) were analyzed in triplicate and the results are shown in Table 35. The formulation containing 100 mg/mL pembrolizumab showed a very low dynamic viscosity of 5.045 mPa*s. The 150 mg/mL pembrolizumab formulation showed an increased but still acceptable viscosity of 19.587 mPa*s, allowing for normal DSP processes, drug substance and drug product filling, and injectability by using standard prefilled syringes for subcutaneous administration.

Table 35: Dynamic viscosity obtained for two different concentrations of pembrolizumab

Example 6: Quantification of sliding yield stress and sliding equilibrium stress by simulating subcutaneous injection of 100 mg/mL and 150 mg/mL pembrolizumab in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/mL polysorbate 80 (pH 5.5 of solution) To test the suitability of the high concentration formulations for subcutaneous administration, 1 mL of both formulations according to Table 27 (containing 100 mg/mL and 150 mg/mL pembrolizumab in 10 mM citric acid/sodium citrate, 205 mM trehalose dihydrate, 0.2 mg/mL polysorbate 80 (pH 5.5 of solution)) were injected into a glass syringe with a nominal capacity of 1 mL with a 29 gauge staked needle (BD Neopak 1MILL 29G TW ¹ / The syringe was filled with ₂₅ mL of 500 mL of 100% 100% 100% 15% 1 ...

After filling, the syringes were tested for injection force simulating injection at an injection rate of 190 mm/min by using a tensile tester (ZwickiLine 500N, ZwickRoell GmbH & Co. KG, Ulm, Germany). The use of the tensile machine allows recording of the applied force in a force-displacement plot. The injection force of the syringe is divided into the sliding yield stress and the sliding equilibrium stress. The sliding yield stress is the force required to initiate the movement of the plunger, and the sliding equilibrium stress (dynamic sliding equilibrium stress) represents the force required to maintain the continuous movement of the plunger.

The plunger rod was attached to the stopper without moving the stopper, and the system was allowed to equilibrate to room temperature before measurements were taken.
Table 36. Injection force of high concentration pembrolizumab loaded into prefilled syringes

The results (see Table 36) show sliding yield stress and sliding equilibrium stress (average and maximum) of less than 15 N for both formulations containing 100 mg/mL and 150 mg/mL pembrolizumab when the syringe contents are expelled. These values allow for easy and safe subcutaneous injection of highly concentrated solutions into patients.

Claims (18)

(a) an anti-human PD1 antibody;
(b) one or more amino acids other than histidine;
(c) a nonionic surfactant;
(d) a citrate buffer,
A liquid pharmaceutical composition, wherein the pH of the composition is 5.0 to 5.8, preferably 5.5. The pharmaceutical composition of claim 1, wherein the citrate buffer is present at a concentration of 1 mM to 50 mM, preferably at a concentration of 5 mM or 10 mM. The pharmaceutical composition according to claim 1 or 2, wherein the non-ionic surfactant is polysorbate 20 or polysorbate 80, preferably polysorbate 80. The pharmaceutical composition according to any one of claims 1 to 3, wherein the non-ionic surfactant is present in a concentration of 0.1 mg/ml to 0.4 mg/ml, preferably 0.2 mg/ml. The pharmaceutical composition according to any one of claims 1 to 4, wherein the amino acid is arginine and/or lysine. The pharmaceutical composition according to any one of claims 1 to 5, comprising arginine at a concentration of 100 mM to 350 mM or 250 mM to 350 mM. The pharmaceutical composition according to any one of claims 1 to 5, comprising lysine at a concentration of 100 mM to 300 mM. The pharmaceutical composition according to any one of claims 1 to 5, comprising arginine and lysine, the total concentration of the arginine and lysine being 100 mM to 350 mM. The pharmaceutical composition according to any one of claims 1 to 8, wherein the anti-human PD1 antibody is pembrolizumab. The pharmaceutical composition according to any one of claims 1 to 9, wherein the anti-human PD1 antibody is present at a concentration of 25 to 80 mg/ml, preferably at a concentration of 25 mg/ml. A liquid pharmaceutical composition comprising a citrate buffer, arginine and/or lysine, polysorbate 20 or polysorbate 80, pembrolizumab and water for injection, and having a pH of 5.0 to 5.8. 12. The liquid pharmaceutical composition of claim 11, comprising: (a) 5 or 10 mM citrate buffer, 300 mM arginine, 0.2 mg/ml polysorbate 80, 25 mg/ml pembrolizumab, and water for injection, and having a pH of 5.0 to 5.8; or (b) 10 mM citrate buffer, 250 mM lysine, 0.2 mg/ml polysorbate 80, 25 mg/ml pembrolizumab, and water for injection, and having a pH of 5.0 to 5.8; or (c) 10 mM citrate buffer, 150 mM arginine, 150 mM lysine, 0.2 mg/ml polysorbate 80, 25 mg/ml pembrolizumab, and water for injection, and having a pH of 5.0 to 5.8. A liquid pharmaceutical composition consisting of 5 mM citrate buffer, 205 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80, 25 mg/ml pembrolizumab and water for injection, having a pH of 5.0 to 5.8, preferably 5.5. histidine, 260-350 mM arginine, polysorbate 20 or polysorbate 80, pembrolizumab and water for injection, having a pH of 5.2-5.8;
Preferably, the liquid pharmaceutical composition consists of 10 mM histidine, 300 mM arginine, 0.2 mg/ml polysorbate 20 or polysorbate 80, 25 mg/ml pembrolizumab and water for injection, and has a pH of 5.2 to 5.8. histidine, sucrose or trehalose dihydrate, polysorbate 20, 90-350 mg/ml pembrolizumab and water for injection, having a pH of 5.2-5.8;
Preferably, a liquid pharmaceutical composition consisting of 10 mM histidine, 140-220 mM trehalose dihydrate, 0.1 mg/ml polysorbate 20, 100 or 150 mg/ml pembrolizumab and water for injection, having a pH of 5.2 to 5.8. A liquid pharmaceutical composition comprising citrate buffer, sucrose or trehalose dihydrate, polysorbate 80, 90-350 mg/ml pembrolizumab, and water for injection, and having a pH of 5.2-5.8. A liquid pharmaceutical composition consisting of 10 mM citrate buffer, 140-220 mM trehalose dihydrate, 0.2 mg/ml polysorbate 80, 100 or 150 mg/ml pembrolizumab, and water for injection, having a pH of 5.0-5.8. 1. A pharmaceutical composition for use in the treatment of cancer, comprising:
The pharmaceutical composition according to any one of claims 1 to 17, wherein the cancer is melanoma, non-small cell lung cancer, classical Hodgkin's lymphoma, urothelial carcinoma, squamous cell carcinoma of the head and neck, renal cell carcinoma, colorectal cancer, small cell lung cancer, microsatellite instability-high or mismatch repair deficient cancer, primary mediastinal large B-cell lymphoma, gastric or gastroesophageal junction adenocarcinoma, hepatocellular carcinoma, Merkel cell carcinoma, endometrial cancer, oncogene hypermutated cancer, cutaneous squamous cell carcinoma, triple-negative breast cancer or cervical cancer.